Novel Phosphate Transporter-B PvPTB1;1/1;2 Contribute to Efficient Phosphate Uptake and Arsenic Accumulation in As-Hyperaccumulator Pteris vittata.

Arsenic (As) contamination in soil poses a potential threat to human health via crop uptake. As-hyperaccumulator Pteris vittata serves as a model plant to study As uptake and associated mechanisms. This study focuses on a novel P/AsV transport system mediated by low-affinity phosphate transporter-B 1 family (PTB1) in P. vittata. Here, we identified two plasma-membrane-localized PTB1 genes, PvPTB1;1/1;2, in vascular plants for the first time, which were 4.4-40-fold greater in expression in P. vittata than in other Pteris ferns. Functional complementation of a yeast P-uptake mutant and enhanced P accumulation in transgenic Arabidopsis thaliana confirmed their role in P uptake. Moreover, the expression of PvPTB1;1/1;2 facilitated the transport and accumulation of As in both yeast and A. thaliana shoots, demonstrating a comparable AsV uptake capacity. Microdissection-qPCR analysis and single-cell transcriptome analysis collectively suggest that PvPTB1;1/1;2 are specifically expressed in the epidermal cells of P. vittata roots. PTB1 may play a pivotal role in efficient P recycling during phytate secretion and hydrolysis in P. vittata roots. In summary, the dual P transport mechanisms consisting of high-affinity Pht1 and low-affinity PTB1 may have contributed to the efficient P/As uptake in P. vittata, thereby contributing to efficient phytoremediation for As-contaminated soils.

Emerging Relevance of Ghrelin in Programmed Cell Death and Its Application in Diseases.

Ghrelin, comprising 28 amino acids, was initially discovered as a hormone that promotes growth hormones. The original focus was on the effects of ghrelin on controlling hunger and satiation. As the research further develops, the research scope of ghrelin has expanded to a wide range of systems and diseases. Nevertheless, the specific mechanisms remain incompletely understood. In recent years, substantial studies have demonstrated that ghrelin has anti-inflammatory, antioxidant, antiapoptotic, and other effects, which could affect the signaling pathways of various kinds of programmed cell death (PCD) in treating diseases. However, the regulatory mechanisms underlying the function of ghrelin in different kinds of PCD have not been thoroughly illuminated. This review describes the relationship between ghrelin and four kinds of PCD (apoptosis, necroptosis, autophagy, and pyroptosis) and then introduces the clinical applications based on the different features of ghrelin.

A Cas12a-based fluorescent microfluidic system for rapid on-site human papillomavirus diagnostics.

Persistent infection with human papillomavirus (HPV) is the leading cause of cervical cancer, and early diagnosis is crucial for clinical management. However, the easy and rapid on-site diagnostic for HPV genotyping remains challenging. Here, we develop a Cas12a-based fluorescent microfluidic detection system for diagnosing six HPV subtypes (HPV6, HPV11, HPV16, HPV18, HPV31, and HPV33). A panel of crRNAs and recombinase polymerase amplification (RPA) primers targeting the HPV L1 gene was screened for sensitive and specific detection. Furthermore, a one-pot RPA reaction was developed to amplify the six HPV subtypes without cross-reactivity. For on-site detection, we integrated the RPA-Cas12a detection into a microfluidic device, enabling the detection of processed clinical samples within 35 minutes. The assay was validated using 112 clinical swab samples and obtained consistent results with the qPCR assay, with a concordance rate of 99.1%. Overall, our diagnostic method offers a rapid, sensitive, and easy-to-use on-site assay for detecting HPV genotypes and holds promise for improving cervical cancer screening and prevention. KEY POINTS: • The Cas12a-based fluorescent microfluidic detection system for the diagnosis of six HPV subtypes. • A one-pot RPA reaction for amplifying the six HPV subtypes without cross-reactivity. • The RPA-Cas12a-microfluidic system provides results within 35 minutes for on-site detection.

Photochemical Transformation of Free Chlorine Induced by Triplet State Dissolved Organic Matter.

Photolysis of free chlorine is an increasingly recognized approach for effectively inactivating microorganisms and eliminating trace organic contaminants. However, the impact of dissolved organic matter (DOM), which is ubiquitous in engineered water systems, on free chlorine photolysis is not yet well understood. In this study, triplet state DOM (3DOM*) was found to cause the decay of free chlorine for the first time. By using laser flash photolysis, the scavenging rate constants of triplet state model photosensitizers by free chlorine at pH 7.0 were determined to be in the range of (0.26-3.33) × 109 M-1 s-1. 3DOM*, acting as a reductant, reacted with free chlorine at an estimated reaction rate constant of 1.22(±0.22) × 109 M-1 s-1 at pH 7.0. This study revealed an overlooked pathway of free chlorine decay during UV irradiation in the presence of DOM. Besides the DOM's light screening ability and scavenging of radicals or free chlorine, 3DOM* played an important role in the decay of free chlorine. This reaction pathway accounted for a significant proportion of the decay of free chlorine, ranging from 23 to 45%, even when DOM concentrations were below 3 mgC L-1 and a free chlorine dose of 70 µM was present during UV irradiation at 254 nm. The generation of HO• and Cl• from the oxidation of 3DOM* by free chlorine was confirmed by electron paramagnetic resonance and quantified by chemical probes. By inputting the newly observed pathway in the kinetics model, the decay of free chlorine in UV254-irradiated DOM solution can be well predicted.

A highly sensitively "off-on-off" fluorescence probe for detection of aluminum ion and water as well as application in Chinese Baijiu.

Developing highly sensitive fluorescent probe for Al3+ and H2O detection is highly desirable, due to aluminum toxicity poses a significant threat to public health. On the other hand, the determination of water content holds immense significance in a wide range of fields such as food processing, pharmaceutical manufacturing. In this paper, a novel acylhydrazone-based fluorescent probe P was successfully synthesized and characterized for the sequential detection of Al3+ and water in alcohols. The probe P exhibited a remarkable "turn-on" response towards Al3+ by emitting yellow fluorescence at 567 nm, with high selectivity and large Stokes shift (147 nm). Meanwhile, the in situ formed P-Al3+ complex demonstrated significant solvatofluorochromic characteristic, which could be utilized as a second probe for detecting water via fluorescence quenching with low detection limit in alcohols (0.008%, methanol; 0.013%, ethanol; 0.013%, isopropanol; 0.037%, n-butanol; vol.%) and acetonitrile (0.072%, vol.%). Moreover, the P-Al3+ complex was able to detect the alcoholic strength of Chinese Baijiu without the interference of other alcohols, providing an excellent recovery rate (100.0-107.0%). Different Chinese Baijius, with various alcoholic strength, could be distinguished by simple test strips. Furthermore, the P-Al3+ complex could also analyze the water content in organic solvents .

Three-in-One Detector by 3D Printing: Simultaneous Contactless Conductivity, Ultraviolet Absorbance, and Laser-Induced Fluorescence Measurements for Capillary Electrophoresis.

We describe a 3-in-1 detector for simultaneous contactless conductivity (C4D), ultraviolet absorbance (UV-AD), and laser-induced fluorescence (LIF) measurements on a single detection point for capillary electrophoresis (CE). A key component of the detector was a rectangular detector head that was assembled with four 3D-printed parts. Two parts covering the detector head to function as a Faraday cage were fused deposition modeling printed using an electrically conductive material. The other two parts in between the conductive parts were stereolithography (SLA) printed with high-resolution (50 µm) constructions on the surface. After assembling the two SLA printed parts, several cavities were built with the surface constructions. Two electrodes and a Faraday shield for C4D were cast by injecting molten Wood's metal into the cavities. For UV-AD, a slit (100 µm width) was created by putting together two grooves (50 µm depth) on the surface of the SLA printed parts. A 255 nm UV-LED was used as the light source. The effective path length and stray light for a 50 µm id capillary were 39 µm and 13%, which were superior to those of other reported 3D-printed AD detectors. Confocal LIF detection was conducted by using an objective lens to focus the laser on the capillary via a through-hole. The detector was used to detect model analytes, including inorganic and organic ions, and fluorescein isothiocyanate labeled amino acids in a signal-run CE separation. In detecting fluorescein, LODs were 1.3 µM (C4D), 2.0 µM (UV-AD), and 1 nM (LIF). The calibration ranges covered from 0.01 µM to 500 µM.

Cinnamaldehyde-modified chitosan hybrid nanoparticles for DOX delivering to produce synergistic anti-tumor effects.

Cancer cells are under oxidative stress associated with the increased generation of reactive oxygen species (ROS). Therefore, increasing the oxidative stress of tumor cells by delivering ROS generators is an effective strategy to induce apoptosis of cancer cells. Herein, we reported a hybrid nanoparticle based on lactobionic acid (LA) modified chitosan and cinnamaldehyde (CA) modified chitosan, which possesses both active tumor-targeting ability and ROS regulation ability, in order to have a synergistic effect with the anti-tumor drug doxorubicin (DOX). LA can improve the tumor-targeting ability and cellular accumulation of these nanoparticles, and CA can induce apoptotic cell death through ROS generation, mitochondrial permeability transition and caspase activation. The particle size and distribution as well as drug release profiles of these nanoparticles were observed. In vitro and in vivo antitumor studies demonstrated that the hybrid nanoparticles show a significant synergistic antitumor effect. Thus, we anticipate that the hybrid nanoparticles have promising potential as an anticancer drug carrier.

Bacillus amyloliquefaciens SC06 alleviates the obesity of ob/ob mice and improves their intestinal microbiota and bile acid metabolism.

Dietary interventions with probiotics have been widely reported to be effective in regulating obesity, and the intestinal microbiota is considered to be an important environmental factor. However, few reports focus on the interactions of microbiota-metabolites-phenotypic variables in ob/ob mice, and they have not been characterized in great detail. In this study, we investigated the effects of Bacillus amyloliquefaciens SC06 on obesity, the intestinal microbiota and the bile acid metabolism of ob/ob mice using biochemical testing, histochemical staining, high-throughput sequencing of the 16S rRNA gene, LC-MS/MS analysis and qRT-PCR. The results showed that SC06 ameliorated the fat mass percentage, hepatic steatosis and liver lipid metabolism disorders and reshaped the gut microbiota and metabolites in male ob/ob mice, specifically deceasing f_S24-7, p_TM7, s_Alistipes massiliensis, f_Rikenellaceae, f_Prevotellaceae, f_Lactobacillaceae, g_Alistipes, g_Flexispira, g_Lactobacillus, g_Odoribacter, g_AF12 and g_Prevotella and increasing f_Bacteroidaceae, g_Bacteroides and f_Desulfovibrionaceae. Meanwhile, SC06 treatment groups had lower ibuprofen and higher glycodeoxycholic acid and 7-dehydrocholesterol. Correlation analysis further clarified the relationships between compositional changes in the microbiota and alterations in the metabolites and phenotypes of ob/ob mice. Moreover, SC06 downregulated bile acid synthesis, export and re-absorption in the liver and increased ileum re-absorption into the blood in ob/ob mice, which may be mediated by the FXR-SHP/FGF15 signaling pathway. These results suggest that Bacillus amyloliquefaciens SC06 can ameliorate obesity in male ob/ob mice by reshaping the intestinal microbial composition, changing metabolites and regulating bile acid metabolism via the FXR signaling pathway.

Protective Effects of Lactobacillus plantarum Lac16 on Clostridium perfringens Infection-Associated Injury in IPEC-J2 Cells.

Clostridium perfringens (C. perfringens) causes intestinal injury through overgrowth and the secretion of multiple toxins, leading to diarrhea and necrotic enteritis in animals, including pigs, chickens, and sheep. This study aimed to investigate the protective effects of Lactobacillus plantarum (L. plantarum) Lac16 on C. perfringens infection-associated injury in intestinal porcine epithelial cell line (IPEC-J2). The results showed that L. plantarum Lac16 significantly inhibited the growth of C. perfringens, which was accompanied by a decrease in pH levels. In addition, L. plantarum Lac16 significantly elevated the mRNA expression levels of host defense peptides (HDPs) in IPEC-J2 cells, decreased the adhesion of C. perfringens to IPEC-J2 cells, and attenuated C. perfringens-induced cellular cytotoxicity and intestinal barrier damage. Furthermore, L. plantarum Lac16 significantly suppressed C. perfringens-induced gene expressions of proinflammatory cytokines and pattern recognition receptors (PRRs) in IPEC-J2 cells. Moreover, L. plantarum Lac16 preincubation effectively inhibited the phosphorylation of p65 caused by C. perfringens infection. Collectively, probiotic L. plantarum Lac16 exerts protective effects against C. perfringens infection-associated injury in IPEC-J2 cells.

Bacillus amyloliquefaciens SC06 Induced AKT-FOXO Signaling Pathway-Mediated Autophagy to Alleviate Oxidative Stress in IPEC-J2 Cells.

Autophagy is a conserved proteolytic mechanism, which degrades and recycles damaged organs and proteins in cells to resist external stress. Probiotics could induce autophagy; however, its underlying molecular mechanisms remain elusive. Our previous study has found that BaSC06 could alleviate oxidative stress by inducing autophagy in rats. This research aimed to verify whether Bacillus amyloliquefaciens SC06 can induce autophagy to alleviate oxidative stress in IPEC-J2 cells, as well as explore its mechanisms. IPEC-J2 cells were first pretreated with 108 CFU/mL BaSC06, and then were induced to oxidative stress by the optimal dose of diquat. The results showed that BaSC06 significantly triggered autophagy, indicated by the up-regulation of LC3 and Beclin1 along with downregulation of p62 in IPEC-J2 cells. Further analysis revealed that BaSC06 inhibited the AKT-FOXO signaling pathway by inhibiting the expression of p-AKT and p-FOXO and inducing the expression of SIRT1, resulting in increasing the transcriptional activity of FOXO3 and gene expression of the ATG5-ATG12 complex to induce autophagy, which alleviated oxidative stress and apoptosis. Taken together, BaSC06 can induce AKT-FOXO-mediated autophagy to alleviate oxidative stress-induced apoptosis and cell damage, thus providing novel theoretical support for probiotics in the prevention and treatment of oxidative damage.

Probiotic Paenibacillus polymyxa 10 and Lactobacillus plantarum 16 enhance growth performance of broilers by improving the intestinal health.

With the ever-growing strict prohibitions on antibiotic growth promoters (AGP) in animal production, in-feed probiotics are becoming attractive alternatives to antibiotics in the poultry industry. To investigate the effects of Paenibacillus polymyxa 10 and Lactobacillus plantarum 16 on the growth performance and intestinal health of broilers, 540 male Cobb 500 broilers of 1 d old were randomly divided into 3 groups with 6 replicates per group and 30 chicks per replicate. Broilers were fed with either a basal diet or basal diets supplemented with 1 × 108 colony-forming units (CFU)/kg P. polymyxa 10 (BSC10) or L. plantarum 16 (Lac16) for 42 d. Results showed that Lac16 treatment improved (P < 0.05) the growth performance (body weight and feed conversion) of broilers at the starter phase, while BSC10 treatment slightly improved (P > 0.05) the growth performance of the starter phase broilers. The increased villus height (P < 0.05) at d 14, 21 and 42 and villus height to crypt depth ratio (P < 0.05) at d 14 and 21 were observed in the ileum of the 2 probiotic groups. Besides, transmission electron microscopy results showed that the 2 probiotics enhanced the intestinal epithelial barrier. Both probiotic treatments up-regulated (P < 0.05) the mRNA expression of fatty acid binding protein 1 (FABP1) and sodium-dependent glucose transporters-1 (SGLT-1) in the ileal mucosa of broilers at d 21. In addition, BSC10 and Lac16 treatments significantly (P < 0.05) increased the relative abundance of short-chain fatty acids-producing bacteria, such as Butyricicoccus pullicaecorum, Faecalibacterium prausnitzii, Lachnospira and Coprococcu, and significantly (P < 0.05) decreased the relative abundance of enteric pathogens (Escherichia coli, Bacteroides fragilis and Shigella sonnei). Furthermore, the 2 probiotic treatments also increased the positive connection among the intestinal microbes and the carbohydrate metabolism-related pathways of the intestinal bacteria (P < 0.05), with decreasing (P < 0.05) nucleotides biosynthesis-related pathways of the intestinal bacteria. Overall, these results suggest that the 2 probiotics, especially Lac16, have a potential beneficial effect on the growth performance and intestinal health of starter phase broilers.

Effects of Bacillus amyloliquefaciens Instead of Antibiotics on Growth Performance, Intestinal Health, and Intestinal Microbiota of Broilers.

The aim of this study was to evaluate the dietary effects of Bacillus amyloliquefaciens SC06 (SC06) instead of antibiotics on the growth performance, intestinal health, and intestinal microbiota of broilers. A total of 360 30-day-old Lingnan yellow broilers were randomly allocated into two groups with six replicates per group (30 birds per replicate). The broilers were fed either a non-supplemented diet or a diet supplemented with 108 colony-forming units lyophilized SC06 per kilogram feed for 30 days. Results showed that SC06 supplementation had no effect on the growth performance compared with that of the control group. SC06 treatment significantly (P <0.05) increased the total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD) activity in the liver, and the activities of trypsin, α-amylase (AMS), and Na+K+-ATPase in the ileum, whereas it decreased (P < 0.05) lipase, gamma glutamyl transpeptidase (γ-GT), and maltase activities in the ileum. Meanwhile, SC06 treatment also improved the immune function indicated by the significantly (P < 0.05) increased anti-inflammatory cytokine [interleukin (IL)-10] level and the decreased (P < 0.05) pro-inflammatory cytokine [IL-6 and tumor necrosis factor (TNF)-α] levels in the ileum. Furthermore, we also found that SC06 enhanced the intestinal epithelial intercellular integrity (tight junction and adhesion belt) in the ileum. Microbial analysis showed that SC06 mainly increased the alpha diversity indices in the jejunum, ileum, and cecum. SC06 treatment also significantly (P < 0.05) increased the abundances of Bacteroidetes, Bacteroidales, Bacteroides, Fusobacteria, Clostridiaceae, and Veillonellaceae in the cecum and simultaneously decreased the abundances of Planococcaceae in the duodenum, Microbacteriaceae in the jejunum, and Lachnospiraceae, [Ruminococcus] and Ruminococcus in cecum. In conclusion, these results suggested that B. amyloliquefaciens instead of antibiotics showed a potential beneficial effect on the intestinal health of broilers.

Effects of Probiotics BaSC06 on Intestinal Digestion and Absorption, Antioxidant Capacity, Microbiota Composition, and Macrophage Polarization in Pigs for Fattening.

This study aimed to compare the effects of BaSC06 and antibiotics on growth, digestive functions, antioxidant capacity, macrophage polarization, and intestinal microbiota of pigs for fattening. A total of 117 pigs for fattening with similar weight and genetic basis were divided into 3 groups: Anti group (containing 40 g/t Kitasamycin in the diet), Anti+Ba group (containing 20 g/t Kitasamycin and 0.5 × 108 CFU/kg BaSC06 in the diet) and Ba group (containing 1 × 108 cfu/Kg BaSC06 in the diet without any antibiotics). Each treatment was performed in three replicates with 13 pigs per replicate. Results showed that BaSC06 replacement significantly improved the ADG (P < 0.05), intestinal digestion and absorption function by increasing the activity of intestinal digestive enzymes and the expression of glucose transporters SGLT1 (P < 0.05) and small peptide transporters PEPT1 (P < 0.05). Besides, BaSC06 supplementation enhanced intestinal and body antioxidant capacity by activating the Nrf2/Keap1 antioxidant signaling pathway due to the increased expression of p-Nrf2 (P < 0.05). Notably, BaSC06 alleviated intestinal inflammation by inhibiting the production of pro-inflammatory cytokines, IL-8, IL-6, and MCP1 (P < 0.05), and simultaneously increasing the expression of M1 macrophage marker protein iNOS (P < 0.05) and M2 macrophage marker protein Arg (P < 0.05) in the intestinal mucosa. Moreover, BaSC06 promoted the polarization of macrophages to M2 phenotype by stimulating the STAT3 signaling pathway. It was also noted that BaSC06 improved microbiota composition by enhancing the proportion of Firmicutes, and reducing that of Bacteroidetes and Proteobacteria. Taken together, our results indicate that dietary supplementation of BaSC06 in pigs for fattening improves the growth, mucosal structure, antioxidative capacity, immune functions (including increasing M1 and M2 polarization of macrophage) and composition of intestinal microbiota, which is much better than antibiotics, suggesting that it is an effective alternative to antibiotics in the preparation of pig feed.

Saccharomyces boulardii attenuates inflammatory response induced by Clostridium perfringens via TLR4/TLR15-MyD8 pathway in HD11 avian macrophages.

Macrophages are professional phagocytic cells that play a critical role in initiating immune responses by presenting antigen and phagocytic clearance. The macrophages can be targeted for immunomodulation by beneficial microbes, such as probiotics. The aim of this study is to investigate the protective effect of Saccharomyces boulardii against Clostridium perfringens infection in avian macrophage cell line HD11. In this study, HD11 macrophages were prestimulated with S. boulardii for 6 h and then infected with C. perfringens for 3 h. Results showed that S. boulardii enhanced phagocytosis and bactericidal capacity against C. perfringens by HD11 cells. The S. boulardii effectively promoted the mRNA expression of CD80, CD83, and CD197 cell-surface molecules in C. perfringens-infected HD11 cells. Moreover, we found that prestimulation with S. boulardii reduced the mRNA expression of CD40, toll-like receptor [TLR] 4, and TLR15 induced by C. perfringens and thereby downregulated the mRNA expression of myeloid differentiation primary response 88, TNF receptor associated factor 6, nuclear factor kappa-B p65 subunit, and c-Jun N-terminal kinase genes in HD11 cells. The upregulation of cytokines (interleukin [IL]-6, tumor necrosis factor alpha, and IL-10) and inducible nitric oxide synthase mRNA expression in C. perfringens-infected HD11 cells were noticeably inhibited by S. boulardii pretreatment. Conclusively, these results might provide a new insight into the role of S. boulardii in regulating avian immune defense against C. perfringens invasion and immune escape.

A Recombinase Polymerase Amplification and Lateral Flow Strip Combined Method That Detects Salmonella enterica Serotype Typhimurium With No Worry of Primer-Dependent Artifacts.

On-site detection demands are quickly increasing to control foodborne pathogenic bacteria along with the long food supply chains. Combining the isothermal recombinase polymerase amplification (RPA) with lateral flow strips (LFSs) is a promising molecular detection approach for the short reaction time, low isothermal condition, and simple and "instrument-free" procedure. However, the method comes with a non-negligible intrinsic risk of the primer-dependent artifacts. In this study, with an important foodborne pathogenic bacterium Salmonella enterica serotype Typhimurium (S. Typhimurium) as the model, system measures including the careful selection of primers targeting unique virulence genes, use of a probe in the RPA reaction, introducing base substitutions with specific guidelines in the primer and probe sequences, and analyzing and screening the primer-probe complex formation were taken to eliminate the primer-dependent artifacts. The measures were strictly tested for the efficacy, and the standardized method was able to specifically detect S. typhimurium within 30 min at 42°C without any interference of probe-primer signals. The established RPA-LFS method shared high sensitivity with the detection limit of 1 CFU/µl of unpurified culture. Our study provided practical measures for the prevention of false positive signals from primer-dimers or primer-probe complexes when using the RPA-LFS method in pathogen detections, and also established a readily applicable method for S. Typhimurium detection.

Application of Bacillus coagulans in Animal Husbandry and Its Underlying Mechanisms.

In recent decades, probiotics have attracted widespread attention and their application in healthcare and animal husbandry has been promising. Among many probiotics, Bacillus coagulans (B. coagulans) has become a key player in the field of probiotics in recent years. It has been demonstrated to be involved in regulating the balance of the intestinal microbiota, promoting metabolism and utilization of nutrients, improving immunity, and more importantly, it also has good industrial properties such as high temperature resistance, acid resistance, bile resistance, and the like. This review highlights the effects of B. coagulans in animal husbandry and its underlying mechanisms.

Protective Effects of Lactobacillus plantarum 16 and Paenibacillus polymyxa 10 Against Clostridium perfringens Infection in Broilers.

This study aimed to investigate the protective effects of Lactobacillus plantarum 16 (Lac16) and Paenibacillus polymyxa 10 (BSC10) against Clostridium perfringens (Cp) infection in broilers. A total of 720 one-day-old chicks were randomly divided into four groups. The control and Cp group were only fed a basal diet, while the two treatment groups received basal diets supplemented with Lac16 (1 × 108 cfu·kg-1) and BSC10 (1 × 108 cfu·kg-1) for 21 days, respectively. On day 1 and days 14 to 20, birds except those in the control group were challenged with 1 × 108 cfu C. perfringens type A strain once a day. The results showed that both Lac16 and BSC10 could ameliorate intestinal structure damage caused by C. perfringens infection. C. perfringens infection induced apoptosis by increasing the expression of Bax and p53 and decreasing Bcl-2 expression and inflammation evidence by higher levels of IFN-γ, IL-6, IL-1ß, iNOS, and IL-10 in the ileum mucosa, and NO production in jejunal mucosa, which was reversed by Lac16 and BSC10 treatment except for IL-1ß (P < 0.05). Besides, the two probiotics restored the intestinal microbiota imbalance induced by C. perfringens infection, characterized by the reduced Firmicutes and Proteobacteria and the increased Bacteroidetes at the phyla level and decreased Bacteroides fragilis and Gallibacterium anatis at the genus level. The two probiotics also reversed metabolic pathways of the microbiota in C. perfringens-infected broilers, including B-vitamin biosynthesis, peptidoglycan biosynthesis, and pyruvate fermentation to acetate and lactate II pathway. In conclusion, Lac16 and BSC10 can effectively protect broilers against C. perfringens infection through improved composition and metabolic pathways of the intestinal microbiota, intestinal structure, inflammation, and anti-apoptosis.

Structural characterization and biological activities of a novel polysaccharide from Phyllanthus emblica.

A novel water-soluble polysaccharide named PEPW80-1, with a molecular mass of 4.7 kDa, was isolated from the pulp tissues of Phyllanthus emblica, and purified by sephadex G-100 column and sephacryl S-300 HR chromatography. The structural features of PEPW80-1 were investigated by a combination of acid hydrolysis, periodate oxidation-Smith degradation, methylation analysis, gas chromatography-mass spectrometry, scanning electron microscope, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. The results showed that PEPW80-1 had a specific optical rotation of [α]25/D = +113° (c = 0.5 mg/mL) and its backbone composed of (1,3)-linked-ß-L-rhamnose and (1,3,6)-linkage-ß-D-galactose, with two branch chains of (1,4)-linked-α-D-galactose and (1,6)-linked-ß-D-galactose and terminated with 1-α-L-arabinose. The antioxidant assays showed that PEPW80-1 possess 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity and hydroxyl radical-scavenging activity, enhancing reductive power. The results of immunomodulatory assays in vitro showed that PEPW80-1 could promote the proliferation of mouse splenocytes. Those proposed that PEPW80-1 might be developed as a potential value-added product with the activities of immunomodulator and free-radical inhibitors.

Inducing effect of dihydroartemisinic acid in the biosynthesis of artemisinins with cultured cells of Artemisia annua by enhancing the expression of genes.

Artemisinin has been used in the production of "artemisinin combination therapies" for the treatment of malaria. Feeding of precursors has been proven to be one of the most effective methods to enhance artemisinin production in plant cultured cells. At the current paper, the biosynthesis of artemisinin (ART) and its four analogs from dihydroartemisinic acid (DHAA) in suspension-cultured cells of Artemisia annua were investigated. ARTs were detected by HPLC/GC-MS and isolated by various chromatography methods. The structures of four DHAA metabolites, namely, dihydro-epi-deoxyarteannuin B, arteannuin I, arteannuin K, and 3-ß-hydroxy-dihydro-epi-deoxyarteannuin B, were elucidated by physicochemical and spectroscopic analyses. The correlation between gene expression and ART content was investigated. The results of RT-PCR showed that DHAA could up-regulate expression of amorpha-4,11-diene synthase gene (ADS), amorpha-4,11-diene C-12 oxidase gene (CYP71AV1), and farnesyl diphosphate synthase gene (FPS) (3.19-, 7.21-, and 2.04-fold higher than those of control group, resp.), which indicated that biosynthesis processes from DHAA to ART were enzyme-mediated.

Stereo and region-selective biosynthesis of two new dihydroartemisinic acid glycosides by suspension-cultured cells of Artemisia annua.

BACKGROUND: The system of plant-cultured cells is one of the optimal systems to investigate biosynthesis pathway and their bioactive intermediates. OBJECTIVE: To study the biosynthesis of dihydroartemisinic acid (1) by suspension-cultured cells of Artemisia annua. MATERIALS AND METHODS: Substrate (compound 1) was administered into the suspension-cultured cells of A. annua and co-cultured for 2 days. The methanol extract was separated on various column chromatography methods and the structures of two biosynthesis products were elucidated based on the analysis of (1)H NMR, (13)C NMR, 2D NMR, and ESI-MS. Time-course curve was also established. Furthermore, in vitro antitumor activities of compounds 1-3 against HepG2, K562, and A549 cell lines were evaluated by MTT assay. RESULTS: Two new compounds were obtained, namely 3α-hydroxy-dihydroartemisinic acid-α-D-glucopyranosyl ester (2) and 15-hydroxy-cadin-4-en-12-oic acid-ß-d-glucopyranosyl ester (3). The results demonstrated that the cultured cells of A. annua possessed the abilities to stereo-selective hydroxylate and region-selective glycosylate sesquiterpene compounds in a highly efficient manner. Inhibitory effects of compounds 1-3 on proliferation of HepG2, K562, and A549 cell lines in vitro were also investigated. CONCLUSION: Two new dihydroartemisinic acid glycosides were obtained by stereo- and region-selective biosynthesis with cultured cells of A. annua.