Transcriptome in Liver of Periparturient Dairy Cows Differs between Supplementation of Rumen-Protected Niacin and Rumen-Protected Nicotinamide.

To investigate the difference between rumen-protected niacin (RPN) and rumen-protected nicotinamide (RPM) in the transcriptome of genes relating to the lipid metabolism of the liver of periparturient dairy cows, 10 healthy Chinese Holstein cows were randomly divided into two groups and fed diets supplemented with 18.4 g/d RPN or 18.7 g/d RPM, respectively. The experiment lasted from 14 days before to 21 days after parturition. Liver biopsies were taken 21 days postpartum for transcriptomic sequencing. In addition, human LO2 cells were cultured in a medium containing 1.6 mmol/L of non-esterified fatty acids and 1 mmol/L niacin (NA) or 2 mmol/L nicotinamide (NAM) to verify the expression of the 10 genes selected from the transcriptomic analysis of the liver biopsies. The expression of a total of 9837 genes was detected in the liver biopsies, among which 1210 differentially expressed genes (DEGs) were identified, with 579 upregulated and 631 downregulated genes. These DEGs were associated mainly with lipid metabolism, oxidative stress, and some inflammatory pathways. Gene ontology (GO) enrichment analysis showed that 355 DEGs were enriched in 38 GO terms. The differences in the expression of these DEGs between RPN and RPM were predominantly related to the processes of steroid catabolism, steroid hydroxylase, monooxygenase activity, oxidoreductase activity, hemoglobin binding, and ferric iron binding, which are involved mainly in lipid anabolism and redox processes. The expressions of FADS2, SLC27A6, ARHGAP24, and THRSP in LO2 cells were significantly higher (p < 0.05) while the expressions of BCO2, MARS1, GARS1, S100A12, AGMO, and OSBPL11 were significantly lower (p < 0.05) on the NA treatment compared to the NAM treatment, indicating that NA played a role in liver metabolism by directly regulating fatty acid anabolism and transport, inflammatory factor expression, and oxidative stress; and NAM functioned more as a precursor of nicotinamide adenine dinucleotide (NAD, coenzyme I) and nicotinamide adenine dinucleotide phosphate (NADP, coenzyme II) to participate indirectly in biological processes such as ether lipid metabolism, cholesterol metabolism, energy metabolism, and other processes.

Effects of Chromium Propionate and Calcium Propionate on Lactation Performance and Rumen Microbiota in Postpartum Heat-Stressed Holstein Dairy Cows.

Chromium propionate (Cr-Pro) and calcium propionate (Ca-Pro) are widely applied in dairy production, especially in the alleviation of heat stress (HS). HS can reduce the abundance of rumen microbiota and the lactation performance of dairy cows. The present work mainly focused on evaluating the effects of Cr-Pro and Ca-Pro on the performance, ruminal bacterial community, and stress of postpartum HS dairy cows as well as identifying the differences in their mechanisms. Fifteen multiparous postpartum Holstein cows with equivalent weights (694 ± 28 kg) and milk yields (41.2 ± 1.21 kg/day) were randomly divided into three groups: control (CON), Cr-Pro (CRPR), and Ca-Pro (CAPR). The control cows received the basal total mixed ration (TMR) diet, while the CRPR group received TMR with 3.13 g/day of Cr-Pro, and the CAPR group received TMR with 200 g/day of Ca-Pro. The rumen microbial 16S rRNA was sequenced using the Illumina NovaSeq platform along with the measurement of ruminal volatile fatty acids (VFAs) and milking performance. Cr-Pro and Ca-Pro improved lactation performance, increased the rumen VFA concentration, and altered the rumen microbiota of the HS dairy cows. Cr-Pro significantly improved the milk yield (p < 0.01). The richness and diversity of the microbial species significantly increased after feeding on Ca-Pro (p < 0.05). Gene function prediction revealed increased metabolic pathways and biological-synthesis-related function in the groups supplemented with Cr-Pro and Ca-Pro. Our results indicate that the application of Cr-Pro or Ca-Pro can provide relief for heat stress in dairy cows through different mechanisms, and a combination of both is recommended for optimal results in production.

Nicotinamide supplementation alters plasma lipidomic profiles of peripartal dairy cows.

Fatty liver syndrome, a common health problem in dairy cows, occurs during the transition from pregnancy to lactation. If the energy supplied to the cow's body cannot meet its needs, a negative energy balance ensues, and the direct response is fat mobilization. Nicotinamide (NAM) has been reported to reduce the nonesterified fatty acid concentration of postpartum plasma. To study the biochemical adaptations underlying this physiologic dysregulation, 12 dairy cows were sequentially assigned to a NAM (45 g/day) treatment or control group. Blood samples were collected on day (D) 1 and D21 relative to parturition. Changes to the plasma lipid metabolism of dairy cows in the two groups were compared using lipidomics. There were significant increases in plasma sphingomyelins d18:1/18:0, d18:1/23:0, d18:1/24:1, d18:1/24:0, and d18:0/24:0 in the NAM group on D1 relative to parturition. In addition, fatty acids 18:2, 18:1, 18:0, 16:1, and 16:0 were obviously decreased on D21 relative to calving. This research has provided insights into how NAM supplementation improves lipid metabolism in perinatal dairy cows.

Fluorination Enhances NIR-II Emission and Photothermal Conversion Efficiency of Phototheranostic Agents for Imaging-Guided Cancer Therapy.

Phototheranostics with second near-infrared (NIR-II) imaging and photothermal effect have become a burgeoning biotechnology for tumor diagnosis and precise treatment. As important parameters of phototheranostic agents (PTAs), fluorescence quantum yield (QY) and photothermal conversion efficiency (PCE) are usually considered as a pair of contradictions that is difficult to be simultaneously enhanced. Herein, a fluorination strategy for designing A-D-A type PTAs with synchronously improved QY and PCE is proposed. Experimental results show that the molar extinction coefficient (ε), NIR-II QY, and PCE of all fluorinated PTAs nanoparticles (NPs) are definitely improved compared with the chlorinated counterparts. Theoretical calculation results demonstrate that fluorination can maximize the electrostatic potential difference by virtue of the high electronegativity of fluorine, which may increase intra/intermolecular D-A interactions, tighten molecule packing, and further promote the increase of ε, ultimately leading to simultaneously enhanced QY and PCE. In these PTA NPs, FY6-NPs display NIR-II emission extended to 1400 nm with the highest NIR-II QY (4.2%) and PCE (80%). These features make FY6-NPs perform well in high-resolution imaging of vasculature and NIR-II imaging-guided photothermal therapy (PTT) of tumors. This study develops a valuable guideline for constructing NIR-II organic PTAs with high performance.

A Facile Structural Isomerization-Induced 3D Spatial D-A Interlocked Network for Achieving NIR-II Phototheranostic Agents.

The performances of second near-infrared (NIR-II) organic phototheranostic agents (OPTAs) depend on both molecular structure and molecular packing when used as nanoparticles (NPs). Herein, we proposed a facile structural isomerization-induced 3D spatial donor (D)-acceptor (A) interlocked network for achieving NIR-II OPTAs. Two isomers, 4MNVDPP and 6MNVDPP were synthesized and formulated into NPs. 6MNVDPP, which has a larger electrostatic potential difference, exhibits a compact 3D spatial D-A interlocked network in the crystal form, while 4MNVDPP forms 2D D-D type J-aggregates. Thus, 6MNVDPP NPs show red-shifted NIR absorption and larger molar extinction coefficient than 4MNVDPP NPs. Thanks to the typical NIR-II emission, superior photothermal-stability, high photothermal conversion efficiency (89 %) and reactive oxygen species production capacity, 6MNVDPP NPs exhibit outstanding NIR-II tiny capillary vasculature/tumor imaging ability and synergistic photothermal/photodynamic anti-cancer effect in vivo.

Exploring the Protective Effects and Mechanism of Huaji Jianpi Decoction against Nonalcoholic Fatty Liver Disease by Network Pharmacology and Experimental Validation.

This paper was designed to predict the mechanisms of the active components of Huaji Jianpi Decoction (HJJPD) against nonalcoholic fatty liver disease (NAFLD) based on network pharmacology-combined animal experiments. The candidate compounds of HJJPD and its relative targets were obtained from TCMSP and PharmMapper web server, and the intersection genes for NAFLD were discerned using OMIM, GeneCards, and DisGeNET. Then, the target protein-protein interaction (PPI) and component-target-pathway networks were constructed. Moreover, gene function annotation (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to study the potential signaling pathways associated with HJJPD's effect on NAFLD. Molecular docking simulation was preformed to validate the binding affinity between potential core components and key targets. Eventually, the candidate targets, the possible pathway, and the mechanism of HJJPD were predicted by the network pharmacology-based strategy, followed by experimental validation in the NAFLD mice model treated with HJJPD. A total of 55 candidate compounds and 36 corresponding genes were identified from HJJPD that are associated with activity against NAFLD, and then the network of them was constructed. Inflammatory response and lipid metabolism-related signaling pathways were identified as the critical signaling pathways mediating the therapeutic effect of the active bioactive ingredients on NAFLD. Compared with the model group, the liver wet weight, liver/body ratio, the levels of total cholesterol (TC), triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and high-density lipoprotein (HDL) in serum in the HJJPD low-dose (17.52 g/kg·d), medium-dose (35.04 g/kg·d), and high-dose (70.07 g/kg·d) groups significantly decreased (P < 0.05). Light microscope observation shows that HJJPD could control the degree of lipid denaturation of the mouse liver tissue to a great extent. RT-qPCR results show that the mRNA expression levels of peroxisome proliferative activated receptor gamma (PPARG), tumor necrosis factor-α (TNF-α), antiserine/threonine protein kinase 1 (AKT1), and prostaglandin-endoperoxide synthase (PTGS2) in the liver tissues of the three HJJPD groups (17.52 g/kg·d, 35.04 g/kg·d, and 70.07 g/kg·d) were significantly lower than those in the model group (P < 0.05). HJJPD can exert its effect by inhibiting hepatic steatosis and related mRNA expression and decreasing the levels of other liver-related indexes. This study suggested that HJJPD exerted its effect on NAFLD by modulating multitargets with multicompounds through multipathways. It also demonstrated that the network pharmacology-based approach might provide insights for understanding the interrelationship between complex diseases and interventions of HJJPD.

Effects of Pamidronate Disodium Combined with Calcium on BMD Values and Severity of Pain in Elderly Patients with Osteoporosis Based on Mobile Terminal Platform for Internet of Things.

Objective: To explore the effects of pamidronate disodium combined with calcium on BMD values and the severity of pain in elderly patients with osteoporosis based on the mobile terminal platform for the Internet of Things. Methods: The data of 120 patients admitted to our hospital from January 2019 to December 2020 were retrospectively analyzed. According to the patients' condition and medication wills, they were divided into the experimental group (n = 68) and the control group (n = 52). All patients were given chronic disease management based on the mobile terminals for the Internet of Things, and they received the treatment of bisphosphonates and calcium, with the supplement of calcium at a daily dose of 1000 mg. The control group was given alendronate sodium once a week, and the experimental group was given pamidronate disodium by intravenous infusion three times a month, with the treatment cycle as 1 year. The patients' bone mineral density (BMD) values and the pain indexes were compared after treatment. Results: There was no statistical difference in general information between the two groups (p > 0.05). The BMD values of the lumbar vertebrae L2-4, total hip, and femur neck at 6 months and 1 year after treatment in the experimental group were significantly higher than those in the control group (p < 0.001). The pain scores at 6 months and 1 year after treatment in the experimental group were significantly lower than those in the control group (p < 0.001). Conclusion: The treatment of pamidronate disodium combined with calcium based on the mobile terminal platform for the Internet of Things can reduce the severity of pain in elderly patients with osteoporosis and improve the BMD, which has a generalization value.

Acceptor Planarization and Donor Rotation: A Facile Strategy for Realizing Synergistic Cancer Phototherapy via Type I PDT and PTT.

Tumor hypoxia seriously impairs the therapeutic outcomes of type II photodynamic therapy (PDT), which is highly dependent upon tissue oxygen concentration. Herein, a facile strategy of acceptor planarization and donor rotation is proposed to design type I photosensitizers (PSs) and photothermal reagents. Acceptor planarization can not only enforce intramolecular charge transfer to redshift NIR absorption but also transfer the type of PSs from type II to type I photochemical pathways. Donor rotation optimizes photothermal conversion efficiency (PCE). Accordingly, three 3,6-divinyl-substituted diketopyrrolopyrrole (DPP) derivatives, 2TPAVDPP, TPATPEVDPP, and 2TPEVDPP, with different number of rotors were prepared. Experimental results showed that three compounds were excellent type I PSs, and the corresponding 2TPEVDPP nanoparticles (NPs) with the most rotors possessed the highest PCE. The photophysical properties of 2TPEVDPP NPs are particularly suitable for in vivo NIR fluorescence imaging-guided synergistic PDT/PTT therapy. The proposed strategy is helpful for exploiting type I phototherapeutic reagents with high efficacy for synergistic PDT and PTT.

Dietary supplementation of hemp oil in teddy dogs: Effect on apparent nutrient digestibility, blood biochemistry and metabolomics.

Present study aimed to evaluate the influence of distinct concentration of dietary supplements hemp oil on apparent nutrient digestibility, blood biochemical parameters and metabolomics of teddy dogs. A total of 25 healthy teddy dogs were selected and divided into five treatments according to diet supplements hemp oil at a rate of 0% (A), 0.5% (B), 1% (C), 2% (D), and 4% (E). Appropriate added hemp oil improved apparent nutrient digestibility of dry matter, crude protein and crude fat (86.32-88.08%, 86.87-88.87% and 96.76-97.43%). The hemp oil significantly increased blood biochemical of utilization related total protein, albumin and globulin (61.33-69.54, 35.08-40.38 and 26.53-31.63 g/L), immunity capacity related immunoglobulin E and γ-interferon (203-347kU/L and 23.04-25.78ng/L), energy-related thyroxine and triiodothyronine (27.11-36.75 and 0.94-1.67 nmol/L). In addition, hemp oil improved superoxide dismutation (26.47-33.02 U/ml) and reduced malondialdehyde (5.30-3.28 nmol/ml). The differential metabolites mainly included nucleotides and metabolites of oxidized lipids, bile and other fatty acids, coenzymes and vitamins. The main metabolic pathways included purine and arachidonic acid metabolism, bile and unsaturated fatty acid biosynthesis, cell oxidative phosphorylation and rheumatoid arthritis. Overall, appropriate dietary supplements hemp oil positively to nutrient digestibility and blood metabolism, immunity and antioxidant capacity, 1% to 2% hemp oil supplements was recommended for teddy dog diet.

Celastrol Loaded Nanoparticles With ROS-Response and ROS-Inducer for the Treatment of Ovarian Cancer.

Ovarian cancer is a gynecological cancer from which it is difficult to be completely cured. It is common to use regimens as an effective treatment for ovarian cancer, but these inevitably bring serious side effects. New treatment strategies and special drugs are needed to improve the prognosis of patients. Celastrol is a natural product, isolated from traditional medicine, that has been proven to be curative for inflammation and cancers. However, the non-targeting and low solubility of celastrol limit its clinical application. We prepared celastrol-loaded nanoparticles for the efficient treatment of ovarian cancer via oxidative stress amplification. In this work, a tumor-targeted, ROS-sensitive nanoparticle was designed, synthesized, and assembled into a drug delivery system that used celastrol. Folic acid (FA) groups on the surface of nanoparticles guide them to actively target the surface of the tumor cell membrane. Thioketal (TK) bonds in nanoparticles can be oxidized and broken into -SH within the ROS level of tumor tissues, which causes the breaking of the PEG hydrophilic shell layer of nanoparticles and promotes the release of celastrol. The released celastrol further stimulated the production of ROS and amplified the intracellular ROS level to promote the apoptosis of tumor cells, thus achieving a therapeutic effect on the celastrol treated ovarian cancer.

Removal kinetics of petroleum hydrocarbons from low-permeable soil by sand mixing and thermal enhancement of soil vapor extraction.

Thermally enhanced remediation of n-alkanes-contaminated silty soil mixed with coarse quartz sands was demonstrated in a laboratory cylindrical tank with diameter of 40 cm and depth of 30 cm. The removal kinetics of semi-volatile n-alkanes (C10, C11, C13-16) under three pulsed heating operations of soil vapor extraction (SVE) was investigated. CMG-STARS software was adopted to simulate the dynamics of heat transfer within the soil column. The results indicated the dramatic increase of air permeability of soil and acceleration of heat transfer after introduction of sand, with the result of achieving rapid soil remediation. Gas-phase transfer of n-alkanes mainly occurred when average soil temperature was ≥100 °C. At the end of remediation with soil subjected to heating for 30.8 h (total running time), the average soil concentration of total n-alkanes was reduced from initial 3106.5 to 202.4 mg/kg, corresponding to 93.4% of mass removal. The residual n-alkanes of C10, C11, C13 and C14 in all collected soil samples were declined to levels of lower than 10 mg/kg. Most of the soil concentration-gradient curves for n-alkanes tested almost coincided with their isothermal contours, indicating the key impact of thermal drive force on contaminant transfer.

Commercial AHAS-inhibiting herbicides are promising drug leads for the treatment of human fungal pathogenic infections.

The increased prevalence of drug-resistant human pathogenic fungal diseases poses a major threat to global human health. Thus, new drugs are urgently required to combat these infections. Here, we demonstrate that acetohydroxyacid synthase (AHAS), the first enzyme in the branched-chain amino acid biosynthesis pathway, is a promising new target for antifungal drug discovery. First, we show that several AHAS inhibitors developed as commercial herbicides are powerful accumulative inhibitors of Candida albicans AHAS (Ki values as low as 800 pM) and have determined high-resolution crystal structures of this enzyme in complex with several of these herbicides. In addition, we have demonstrated that chlorimuron ethyl (CE), a member of the sulfonylurea herbicide family, has potent antifungal activity against five different Candida species and Cryptococcus neoformans (with minimum inhibitory concentration, 50% values as low as 7 nM). Furthermore, in these assays, we have shown CE and itraconazole (a P450 inhibitor) can act synergistically to further improve potency. Finally, we show in Candida albicans-infected mice that CE is highly effective in clearing pathogenic fungal burden in the lungs, liver, and spleen, thus reducing overall mortality rates. Therefore, in view of their low toxicity to human cells, AHAS inhibitors represent a new class of antifungal drug candidates.

A regulatory mutant on TRIM26 conferring the risk of nasopharyngeal carcinoma by inducing low immune response.

The major histocompatibility complex (MHC) is most closely associated with nasopharyngeal carcinoma (NPC), but the complexity of its genome structure has proven challenging for the discovery of causal MHC loci or genes. We conducted a targeted MHC sequencing in 40 Cantonese NPC patients followed by a two-stage replication in 1065 NPC cases and 2137 controls of Southern Chinese descendent. Quantitative RT-PCR analysis (qRT-PCR) was used to detect gene expression status in 108 NPC and 43 noncancerous nasopharyngeal (NP) samples. Luciferase reporter assay and chromatin immunoprecipitation (ChIP) were used to assess the transcription factor binding site. We discovered that a novel SNP rs117565607_A at TRIM26 displayed the strongest association (OR = 1.909, Pcombined = 2.750 × 10-19 ). We also observed that TRIM26 was significantly downregulated in NPC tissue samples with genotype AA/AT than TT. Immunohistochemistry (IHC) test also found the TRIM26 protein expression in NPC tissue samples with the genotype AA/AT was lower than TT. According to computational prediction, rs117565607 locus was a binding site for the transcription factor Yin Yang 1 (YY1). We observed that the luciferase activity of YY1 which is binding to the A allele of rs117565607 was suppressed. ChIP data showed that YY1 was binding with T not A allele. Significance analysis of microarray suggested that TRIM26 downregulation was related to low immune response in NPC. We have identified a novel gene TRIM26 and a novel SNP rs117565607_A associated with NPC risk by regulating transcriptional process and established a new functional link between TRIM26 downregulation and low immune response in NPC.

A multifunctional luminogen with aggregation-induced emission characteristics for selective imaging and photodynamic killing of both cancer cells and Gram-positive bacteria.

The increasing impact of bacteria on cancer progression and treatments has been witnessed in recent years. Insufficient attention to cancer-related bacteria may lead to distant metastasis, poor therapeutic efficiency and low survival rates for cancers. Exploiting new approaches that enable selective imaging and effective killing of cancer cells and bacteria are thus of great value for the battle against cancers. Herein, we report an aggregation-induced emission (AIE) luminogen, namely TPPCN, with intense emission and efficient reactive oxygen species production for fluorescence imaging and killing cancer cells and Gram-positive bacteria. This work not only demonstrates the potential of AIE luminogens in comprehensive cancer treatments but also stimulates the enthusiasm of scientists to design more multifunctional AIE systems for both cancer and bacteria theranostics.

Madecassoside suppresses proliferation and invasiveness of HGF-induced human hepatocellular carcinoma cells via PKC-cMET-ERK1/2-COX-2-PGE2 pathway.

Recent studies showed that Madecassoside (MAD), a pentacyclic triterpene isolated from Centella asitica (L.), was used as a therapeutic agent in wound healing and also as an anti-inflammatory, anti-oxidative activities and anti-aging agent. However, its role in cancer has not been elucidated. In our present study, hepatocyte growth factor (HGF) induced the phosphorylation of its corresponding receptor cMET, increased expression of cyclo-oxygenase-2 (COX-2) and prostaglandin E2 (PGE2) in human hepatocellular carcinoma (HCC) cells lines (HepG2 and SMMC-77), and this effect was inhibited by MAD in a dose-dependent manner. In addition, MAD exhibited significant anti-proliferative and anti-invasive effect in HGF-induced HepG2 and SMMC-77 cells. Moreover, MAD inhibited the phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) and the protein kinase C (PKC) activity in HGF-induced HepG2 and SMMC-77 cells. This conclusion was consistent with the effect of selective COX-2 inhibitor (NS-398) and knockdown of COX-2 by siRNA on attenuating the proliferation and invasiveness potential, and over-expression of COX-2 on abolishing the effects of MAD on proliferation and invasiveness potential, and was also in parallel with the effect of PKC inhibitor (Bisindolylmaleimide) on inhibiting PKC activity, MEK/ERK1/2 inhibitor (PD98059) inhibited MEK/ERK1/2 pathways in HGF-induced HepG2 and SMMC-77 cells. Collectively, MAD could inhibit the HGF-activated proliferation and invasiveness of HCC cells via regulating the activation of cMET-PKC-ERK1/2-COX-2-PGE2 cascade, which indicated that MAD might help control HGF-linked HCC.

Influence of Dietary Copper on Serum Growth-Related Hormone Levels and Growth Performance of Weanling Pigs.

To investigate the effect of dietary copper on serum growth-related hormones levels and growth performance, a total of 60 weanling pigs were randomly assigned to six groups each containing 10 pigs, fed on basal diets supplemented with 0 (control), 100, 150, 200, 250, and 300 mg/kg copper sulfate for 80 days, respectively. The average daily gain (ADG), feed to gain ratio (F/G), feed intake and serum growth hormone (GH), insulin (INS), insulin-like growth factor 1 (IGF-1), and insulin-like growth factor-binding protein 3 (IGFBP-3) levels were detected at interval of 20 days. The results revealed that ADG, and serum GH, INS, IGF-1, and IGFBP-3 concentrations were increased significantly in the pigs fed on diets added with 100, 150, 200, 250, and 300 mg/kg copper sulfate. Meanwhile, in the pigs supplemented with 250 mg/kg copper sulfate, ADG was increased significantly from the 40th to the 60th day of the experiment (P < 0.01), and the levels of GH, INS, IGF-1, and IGFBP-3 in serum were elevated significantly from the 20th to the 40th day of the experiment (P < 0.01). It is concluded that effects of copper supplemented in the diet on the growth of pigs were related to the increasing levels of GH, INS, IGF-1, and IGFBP-3 in serum which were induced by copper. High dietary copper increase the concentrations of growth-related hormones in serum, resulting in improving the growth performance of weanling pigs.

Characterization and immunomodulatory activities of polysaccharides from Spirogyra neglecta (Hassall) Kützing.

Sulfated polysaccharides (SP) isolated from freshwater green algae, Spirogyra neglecta (Hassall) Kützing, and fractionated SPs were examined to investigate their molecular characteristics and immunomodulatory activity. The crude and fractionated SPs (F1, F2, and F3) consisted mostly of carbohydrates (68.5-85.3%), uronic acids (3.2-4.9%), and sulfates (2.2-12.2%) with various amounts of proteins (2.6-17.1%). D-galactose (23.5-27.3%), D-glucose (11.5-24.8%), L-fucose (19.0-26.7%), and L-rhamnose (16.4-18.3%) were the major monosaccharide units of these SPs with different levels of L-arabinose (3.0-9.4%), D-xylose (4.6-9.8%), and D-mannose (0.4-2.3%). The SPs contained two sub-fractions with molecular weights (Mw) ranging from 164 × 10(3) to 1460 × 10(3) g/mol. The crude and fractionated SPs strongly stimulated murine macrophages, producing considerable amounts of nitric oxide and various cytokines via up-regulation of their mRNA expression by activation of nuclear factor-kappa B and mitogen-activated protein kinases pathways. The main backbone of the most immunoenhancing SP was (1→3)-L-Fucopyranoside, (1→4,6)-D-Glucopyranoside, and (1→4)-D-Galactopyranoside.

Simultaneous determination of 15 phenolic constituents of Chinese black rice wine by HPLC-MS/MS with SPE.

UNLABELLED: This study established a new method for quantitative and qualitative determination of certain components in black rice wine, a traditional Chinese brewed wine. Specifically, we combined solid-phase extraction and high-performance liquid chromatography (HPLC) with triple quadrupole mass spectrometry (MS/MS) to determine 8 phenolic acids, 3 flavonols, and 4 anthocyanins in black rice wine. First, we clean samples with OASIS HLB cartridges and optimized extraction parameters. Next, we performed separation on a SHIM-PACK XR-ODS column (I.D. 3.0 mm × 75 mm, 2.2 µm particle size) with a gradient elution of 50% aqueous acetonitrile (V/V) and water, both containing 0.2% formic acid. We used multiple-reaction monitoring scanning for quantification, with switching electrospray ion source polarity between positive and negative modes in a single chromatographic run. We detected 15 phenolic compounds properly within 38 min under optimized conditions. Limits of detection ranged from 0.008 to 0.030 mg/L, and average recoveries ranged from 60.8 to 103.1% with relative standard deviation ≤8.6%. We validated the method and found it to be sensitive and reliable for quantifying phenolic compounds in rice wine matrices. PRACTICAL APPLICATION: This study developed a new, reliable HPLC-MS/MS method for simultaneous determination of 15 bioactive components in black rice wine. This method was validated and found to be sensitive and reliable for quantifying phenolic compounds in rice wine.

[A research on the influence of two herbal concoctions on Toll-like receptor signal pathways of influenza virus induced pneumonia in mice].

OBJECTIVE: To investigate the effect of Shufeng Xuanfei and Jiebiao Qingli concoctions on Toll-like receptor (TLR) signal pathway of pneumonia infected with influenza virus in mice. METHODS: The pneumonia model was reproduced by nasal dropping of influenza virus A in mice. The mice were randomly divided into nine groups: normal group (C), model group (M), tamiflu group (D), Shufeng Xuanfei low-dose (SL), medium-dose (SM) and high-dose (SH) groups, Jiebiao Qingli low-dose (JL), medium-dose (JM) and high-dose (JH) groups, each n=12. Two hours after model-reproduction, the mice in C group and M group received distilled water by gavage. The mice in D group received 2.5 g×mL(-1)×d(-1) oseltamivir phosphate. Shufeng Xuanfei formula in doses of 3.76, 1.88, 0.94 g×kg(-1)×d(-1) were respectively administered to SH, SM and SL groups by gavage, Jiebiao Qingli formula in doses of 4.37, 2.18, 1.09 g×kg(-1)×d(-1) was given to JH, JM and JL groups by gavage, respectively. Each group was in equal dose of 0.2 mL daily over a 4-day period. Total RNA was extracted in each group. Then gene chips were used to screen these RNA samples. Some genes that were involved in TLR signal pathways were selected. These candidate genes were verified by real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: TLR7, MYD88, CCL5, IFNB1, IL6, IL12a, NFKBIA and IKBKB were up-regulated in model group compared with control group. Compared with model group, down-regulated genes in medium-dose, low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula included TLR3, TLR7, MYD88, CCL5, IFNB1, IL6, IL12a, NFKBIA and IKBKB (log2 signal intensity of SM/M in medium-dose Shufeng Xuanfei formula group were -1.24, -2.02, -1.36, -1.95, -0.63, -1.33, -3.50, -1.33, -1.33, log2 signal intensity of SL/M in low-dose Shufeng Xuanfei group were -1.07, -2.43, -2.63, -2.30, -5.09, -3.19, -3.53, -1.95, -1.95, log2 signal intensity of JM/M in medium-dose Jiebiao Qingli formula group were -1.78, -0.55, -1.35, -1.47, -1.65, -2.03, -3.02, -1.57, -1.57, respectively). The results suggested that the effect of Shufeng Xuanfei formula was better than that of Jiebiao Qingli formula. By RT-PCR, compared with model group, low-dose, medium-dose and high-dose groups of Shufeng Xuanfei formula, medium-dose and high-dose groups of Jiebiao Qingli formula, and tamiflu group, significant decrease in TLR7, nuclear factor-ΚB (NF-ΚB), myeloid differential protein-88 (MyD88) mRNA expression were found. Medium-dose and low-dose Shufeng Xuanfei formula group (TLR7 mRNA: 3.6±0.3, 3.5±1.2 vs. 7.4±1.6, NF-ΚB mRNA: 1.1±0.2, 1.0±0.2 vs. 2.2±0.4; MyD88 mRNA: 1.4±0.4, 1.0±0.3 vs. 3.4±0.9, all P<0.01) and medium-dose Jiebiao Qingli formula group (TLR7 mRNA: 4.9±0.3 vs. 7.4±1.6, NF-ΚB mRNA: 1.3±0.7 vs. 2.2±0.4, MyD88 mRNA: 1.6±0.8 vs. 3.4±0.9, P<0.05 or P<0.01) were shown statistically significant decreases compared with the model group. CONCLUSIONS: Medium-dose and low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula can inhibit the inflammatory reaction induced by influenza virus by down-regulating the NF-ΚB through TLR signal pathways dependent on MyD88. The regulation of Shufeng Xuanfei formula in TLR signal pathways was superior to that of Jiebiao Qingli formula.

Immunomodulatory of selenium nano-particles decorated by sulfated Ganoderma lucidum polysaccharides.

In this study, we employed a one-step method to prepare selenium nanoparticles (SeNPs) decorated by the water-soluble derivative of Ganoderma lucidum polysaccharides (SPS). The SeNPs-SPS complexes were stable, and the diameter of the SeNPs was homogeneous at around 25 nm. We investigated the anti-inflammatory activity of SeNPs-SPS against murine Raw 264.7 macrophage cells induced by LPS. SeNPs-SPS were found to significantly inhibit LPS-stimulated nitric oxide (NO) production against Raw 264.7 macrophages. RT-PCR results reveal the down-regulation of mRNA gene expressions for pro-inflammatory cytokines, including inducible NO synthase (iNOS), interleukin (IL)-1 and TNF-α in a dose-dependent manner. However, the anti-inflammation cytokine IL-10 was markedly increased. In the NF-κB signal pathway, SeNPs-SPS significantly inhibited the phosphorylation of Iκ-Bα. Similar results were observed for inhibition of the phosphorylation of JNK1/2 and p38 mitogen-activated protein kinase(MAPKs), whereas ERK1/2 MAPK was not apparently affected by SeNPs-SPS. All of these results suggest that SeNPs-SPS complexes have anti-inflammatory potential modulating pro-/anti-inflammation cytokine secretion profiles, and that the mechanism is partially due to inhibition of activations of NF-κB, JNK1/2 and p38 MAPKs.