Protective Mechanism of Polysaccharide ORP-1 Isolated from Oudemansiella raphanipes against Age-Related Cognitive Decline through the Microbiota-Gut-Brain Axis.

Age-related cognitive decline is primarily attributed to the progressive weakening of synaptic function and loss of synapses, while age-related gut microbial dysbiosis is known to impair synaptic plasticity and cognitive behavior by metabolic alterations. To improve the health of the elderly, the protective mechanisms of Oudemansiella raphanipes polysaccharide (ORP-1) against age-related cognitive decline are investigated. The results demonstrate that ORP-1 and its gut microbiota-derived metabolites SCFAs restore a healthy gut microbial population to handle age-related gut microbiota dysbiosis mainly by increasing the abundance of beneficial bacteria Dubosiella, Clostridiales, and Prevotellaceae and reducing the abundance of harmful bacteria Desulfovibrio, strengthen intestinal barrier integrity by abolishing age-related alterations of tight junction (TJ) and mucin 2 (MUC2) proteins expression, diminish age-dependent increase in circulating inflammatory factors, ameliorate cognitive decline by reversing memory- and synaptic plasticity-related proteins levels, and restrain hyperactivation of microglia-mediated synapse engulfment and neuroinflammation. These findings expand the understanding of prebiotic-microbiota-host interactions.

Polysaccharide ORP-1 isolated from Oudemansiella raphanipes ameliorates age-associated intestinal epithelial barrier dysfunction in Caco-2 cells monolayer.

Age-associated increase in intestinal permeability is known to relate with gut microbiota dysbiosis and loss of epithelial tissue integrity. To improve healthy aging and prevent age-associated chronic disabilities, the protective potential of polysaccharides from Oudemansiella raphanipes (ORP-1) against age-associated intestinal epithelial barrier dysfunction in d-galactose-induced Caco-2 cells monolayer was investigated. In-vitro results demonstrated that ORP-1 can restore a healthy gut microbial population to handle age-related gut microbiota dysbiosis mainly by facilitating the proliferation and adhesion of probiotics Lactobacillus acidophilus (L. acidophilus) and Bifidobacterium bifidum (B. bifidum) to compete with intestinal pathogenic Escherichia coli (E. coli) for ecological niches and nutrition. Meanwhile, ORP-1 strengthened the intestinal structural integrity primarily by abolishing the aggravation of apoptosis and the age-associated alterations of tight junction (TJ) proteins expression in intestine. These findings highlighted that ORP-1 could be a potential functional food component with preventive utility against age-associated intestinal barrier injury.

Reducing tumescent anesthetic injection pain by topical anesthesia pretreatment among patients undergoing endovenous radiofrequency ablation of varicose veins: A double-blind randomized controlled trial.

OBJECTIVES: Tumescent anesthesia frequently causes the intraoperative and postoperative pain during radiofrequency ablation (RFA) of varicose veins. We have to find a way to reduce pain caused by these injections. This randomized controlled trial investigated the effectiveness of topical anesthesia pretreatment (TAP) on relieving needle puncture pain during administration of tumescent anesthesia among patients undergoing RFA of varicose veins. METHODS: Eligible patients treated with RFA were recruited and randomized to either application of TAP with lidocaine-prilocaine cream (EMLA) or water-based cream (placebo). The primary outcome was patient described pain scores on the visual analogue scale (VAS) at different time points during the procedure. Secondary outcomes were technical success rate, complications, satisfaction level, expense, and extra analgesia use. RESULTS: Sixty-two patients were randomized: 32 to EMLA and 30 to placebo. Both groups had comparable baseline demographics, CEAP classification, and Venous Clinical Severity Score (VCSS). Less tumescent anesthetic needle puncture pain was found in the EMLA group (22 ± 7 vs 42 ± 8, p < .01). Pain scores of other time points were equivalent. There was less pain in EMLA pretreated area compared to non-pretreated area in the same patient during needle puncture (22 ± 7 vs 45 ± 7, p < .01), and similar phenomena did not appear in the placebo group. There was no statistical difference in complications, satisfaction level, expense, and technical success between the two groups. And no extra analgesia was used in all patients. CONCLUSION: We recommend the routine use of TAP to reduce the needle puncture pain during tumescent anesthesia in RFA of lower extremity varicose veins.

Characterization and immunomodulatory effect of an alkali-extracted galactomannan from Morchella esculenta.

In our continuous exploration for bioactive polysaccharides, a novel polysaccharide FMP-2 was isolated and purified from the fruiting bodies of Morchella esculenta by alkali-assisted extraction. FMP-2 had an average molecular weight of 1.09 × 106 Da and contained mannose, glucuronic acid, glucose, galactose, and arabinose in a molar ratio of 4.10:0.22:1.00:5.75:0.44. The backbone of FMP-2 mainly consisted of 1,2-α-D-Galp, 1,6-α-D-Galp, and 1,4-α-D-Manp, with branches of 1,4,6-α-D-Manp and 1,2,6-α-D-Galp. FMP-2 can stimulate phagocytosis and promote the secretion of NO, ROS, and cytokines like IL-6, IL-1ß, and TNF-α in RAW264.7 cells ranging from 25 to 400 µg/mL. FMP-2 had great repairing effect on the immune injury of zebrafish induced by chloramphenicol. The phagocytosis ability of zebrafish macrophages and the proliferation of neutrophils can be greatly enhanced by polysaccharide FMP-2 with concentrations from 50 to 200 µg/mL. These findings suggest that FMP-2 might be used as a potential immunomodulator in the food and pharmaceutical industries.

Efficacy and safety of DOACs in patients with peripheral arterial disease: A systematic review and meta-analysis.

BACKGROUND: The efficacy and safety of direct oral anticoagulants (DOACs) in patients with peripheral arterial disease are not completely understood. Therefore, we conducted a meta-analysis to explore the effects of DOACs in this population. METHODS: We systematically searched the PubMed, Cochrane Library, and Web of Science till April 2020 for relevant randomized controlled trials and observational studies, with no linguistic restrictions. The efficacy outcomes were cardiovascular death, stroke, myocardial infraction, major adverse cardiovascular events (MACE), acute limb ischemia, amputation, and target lesion revascularization. The safety outcome was major bleeding events. Random effects risk ratios with 95% confidence intervals were calculated. RESULTS: A total four randomized controlled trials were included in this meta-analysis. Among peripheral arterial disease patients, DOACs did not reduce the risk of cardiovascular death (RR = 1.02 95%CI 0.75-1.37, P = 0.92), stroke (RR = 0.73 95%CI 0.46-1.14, P = 0.16), myocardial infraction (RR = 0.85 95%CI 0.70-1.03, P = 0.10), MACE (RR = 0.73 95%CI 0.46-1.14, P = 0.16), or amputation (RR = 0.73 95%CI 0.46-1.14, P = 0.16) compared with control. However, DOACs were associated with reduction in acute limb ischemia (RR = 0.67 95%CI 0.55-0.80, P < 0.01) and target lesion revascularization (RR = 0.89 95%CI 0.81-0.99, P = 0.02) at the expense of major bleeding events (RR = 1.43 95%CI 1.16-1.77, P < 0.01) compared with control. CONCLUSIONS: Based on current evidence, no significant difference in cardiovascular death, stroke, myocardial infraction, MACE, and amputation was found when DOACs were compared to antiplatelet monotherapy. The benefits of preventing target lesion revascularization and acute limb ischemia were balanced by amplified risk of major bleeding. Larger randomized controlled trials are needed to figure out the uncertainty around efficacy and safety of medications for peripheral arterial disease.

Reduction of 5-fluorouracil-induced toxicity by Sarcodon aspratus polysaccharides in Lewis tumor-bearing mice.

5-Fluorouracil (5-Fu) is an effective anticarcinogenic agent, however, continuous use of 5-Fu may cause severe side effects. The goal of this study was to investigate the effectiveness of Sarcodon aspratus polysaccharides (SATP) in alleviating 5-Fu-induced toxicity in Lewis tumor-bearing mice. Lewis tumor-bearing mice were treated with saline, SATP, 5-Fu or 5-Fu + SATP. The results indicated that compared to the 5-Fu group, the 5-Fu + SATP group showed effective amelioration of the liver, kidney and small intestine injury caused by 5-Fu and decreases in the levels of related biochemical indicators, such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) and urea nitrogen (BUN). Additionally, the combination therapy enhanced the quality of life and immune organ indexes of mice. Further mechanistic studies indicated that the 5-Fu + SATP group showed a decrease in hepatotoxicity caused by 5-Fu via a reduction in the levels of interleukin-1ß (IL-1ß), an increase in the expression of Bcl-2 and decreases in the expression of p-p38, p-JNK and Bax. Collectively, the results indicated that SATP could significantly alleviate the toxicity of 5-Fu in Lewis tumor-bearing mice and showed the hepatoprotective capability of SATP via its effect on the expression levels of inflammatory factors and components of the MAPK/P38/JNK pathway, which shows that it may be a potential adjuvant for the chemotherapeutic drug 5-Fu in cancer treatment.

Structural characterization and in vitro hypoglycemic activity of a glucan from Euryale ferox Salisb. seeds.

In this research, a polysaccharide fraction (EFSP-1) was obtained from the seeds of Euryale ferox Salisb. by DEAE sepharose FF and Superdex™ 75 gel chromatography. The average molecular weight (Mw) of EFSP-1 was 8.75 kDa. Monosaccharides composition analysis indicated that EFSP-1 was a glucan. The structure of EFSP-1 was characterized by analysis of FT-IR, GC-MS and NMR, which indicated that the backbone of EFSP-1 was mainly composed of (1→4)-α-D-Glcp with branches substituted at O-6 and terminated with T-α-D-Glcp. Moreover, the hypoglycemic effect of EFSP-1 was investigated by establishing insulin resistance HepG2 and 3T3-L1 cells. The results showed that EFSP-1 could increase glucose consumption by up-regulating the expression of GLUT-4 via activating PI3K/Akt signal pathway in IR cells. Hence, EFSP-1 could be a potential functional food to ameliorate insulin resistance for diabetes therapy.

Immunomodulatory effect of a polysaccharide fraction on RAW 264.7 macrophages extracted from the wild Lactarius deliciosus.

The mushroom polysaccharides are important substances with variety of functions, especially to the human body's immunomodulation effects. In this work, a polysaccharide fraction (LDP-1) was extracted and purified from the fruiting bodies of a rare wild Lactarius deliciosus. LDP-1 with molecular weight of 9.8 × 105 Da showed an obvious immunological activity to the RAW 264.7 cells. It had no significant suppressive but promotive effects on proliferation of the macrophages. The production of nitric oxide (NO) presented a concentration-dependent manner after treated with the LDP-1, and the maximum yield of NO was 39.15 µM. LDP-1 could promote the phagocytic uptake ability of the RAW 264.7 cells significantly, and many of the antennas produced around the cells correspondingly. The cytokines of TNF-α, IL-1ß and IL-6 were secreted increasingly in a concentration-dependent manner, which were 4.83, 17.8 and 11 times than that of the control, respectively. Western blotting analysis confirmed that NF-κB levels in the nucleus were increased while cytoplasmic inhibitor of NF-κB (IκB-α) degraded after treated with the LDP-1, indicating the RAW 264.7 cells probably be stimulated by LDP-1 through activating the IκB-α-NF-κB pathway. These results demonstrated that LDP-1 could be used as a kind of immunomodulatory agent for healthcare potentially.

Anti-inflammatory effects of Morchella esculenta polysaccharide and its derivatives in fine particulate matter-treated NR8383 cells.

Fine particulate matter (PM2.5) exposure could cause many acute and chronic respiratory diseases. In this study the protective effects of polysaccharide from Morchella esculenta (FMP-1) and its derivatives against PM2.5-induced inflammation were evaluated. By flow cytometry and ELISA analysis, sulfated polysaccharide SFMP-1 showed the best protective effect in reducing PM2.5-induced cell death, cell apoptosis and production of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1ß), which was accompanied by a diminished level in reactive oxygen species (ROS) formation caused by PM2.5 in rat alveolar macrophage NR8383 cells. Furthermore, the mechanism was studied by immunofluorescence, qRT-PCR and western blotting. SFMP-1 could down-regulate the expression of inducible NO synthesis (iNOS) and cyclooxygenase-2 (COX-2) at both mRNA and protein levels in PM2.5-treated cells. The PM2.5-induced phosphorylation of nuclear factor-kappa B (NF-κB) was also reduced through suppressing nuclear translation of the NF-κB and inhibiting the degradation and phosphorylation of IκBα. These results indicated that SFMP-1 could protect NR8383 cells from PM2.5-induced inflammation by inhibiting NF-κB activation.

Effect of polysaccharide FMP-1 from Morchella esculenta on melanogenesis in B16F10 cells and zebrafish.

Polysaccharides from Morchella esculenta are known to exhibit diverse bioactivities, while an anti-melanogenesis effect has been barely addressed. Herein, the anti-melanogenesis activity of a heteropolysaccharide from M. esculenta (FMP-1) was investigated in vitro and in vivo. FMP-1 had no significant cytotoxic effect on B16F10 melanoma cells as well as zebrafish larvae, but did reduce melanin contents and tyrosinase activities in both of them. Treatment with FMP-1 also effectively suppressed the expression of melanogenesis-related proteins, including MC1R, MITF, TRP-1 and TRP-2, through decreasing the phosphorylation of cyclic adenosine monophosphate response element-binding protein (CREB). Moreover, the mitogen-activated protein kinase (MAPK) pathway was observed mediating FMP-1's inhibitory effect against melanin production. Specifically, FMP-1 treatment markedly inhibited the activation of phosphorylation of p38 mitogen-activated protein kinase. These results suggested that FMP-1's inhibitory effect against melanogenesis is mediated by the inhibition of CREB and p38 signaling pathways, thereby resulting in the downstream repression of melanogenesis-related proteins and the subsequent melanin production. These data provide insight into FMP-1's potential anti-melanogenesis effect in food and cosmetic industries.

Polysaccharide FMP-1 from Morchella esculenta attenuates cellular oxidative damage in human alveolar epithelial A549 cells through PI3K/AKT/Nrf2/HO-1 pathway.

Oxidative stress is considered to involve cell death in severe pulmonary diseases like idiopathic pulmonary fibrosis (IPF). Polysaccharide FMP-1 from Morchella esculenta can exert significant antioxidant activity. However, its effects on alveolar epithelial cells remain unperceived. Herein, the effects of FMP-1 against H2O2-induced oxidative damage in human alveolar epithelial A549 cells were investigated. FMP-1 could inhibit H2O2-induced cytochrome C and Caspase-3 release to prevent cell apoptosis via attenuation of MDA and ROS levels, and enhancement the enzymatic activities of SOD and T-AOC. Furthermore, the underlying molecular mechanisms were clarified. The phosphorylation of AKT and the nuclear translocation of Nrf2 were observed to be promoted by FMP-1 as well as the level of HO-1. These findings suggested that FMP-1 attenuate cellular oxidative stress through PI3K/AKT pathway, and FMP-1 could be explored as natural potential antioxidants to lower oxidative stress relevant to the progression of IPF.

Structural characterization and antioxidant activities of a novel polysaccharide fraction from the fruiting bodies of Craterellus cornucopioides.

A new polysaccharide fraction (CCPP-1) was obtained from Craterellus cornucopioides. CCPP-1 had an average molecular weight of 9.2 × 105 Da, which was mainly composed of mannose, glucose, xylose, arabinose, fructose in molar ratio of 0.7:0.05:0.18:1:0.05. Results of structural characterization revealed that the dominant linkage types of CCPP-1 were →3, 6)-Manp(1→, T-Araf, →4, 6)-Manp (1→, →5)-Araf (1→ and →3)-Araf (1→. Interesting, in vitro antioxidant activities assays showed that CCPP-1 possessed strong scavenging abilities on DPPH and ABTS radicals. The oxidative hemolysis induced by AAPH in mice erythrocytes was effectively reversed by incubation with CCPP-1. CCPP-1 significantly prevented AAPH-induced intracellular reactive oxygen species (ROS) generation. Moreover, CCPP-1 could significantly restore AAPH-induced increase of intracellular antioxidant enzyme glutathione peroxidase (GPx) and catalase (CAT) activities to normal level, as well as inhibit intracellular malondialdehyde (MDA) formation. Therefore, CCPP-1 could protect against AAPH-induced oxidative-stress in erythrocytes, which would be explored as naturally potential antioxidant agent applied in food and cosmetic fields.

Structural characterization, in vitro and in vivo antioxidant activities of a heteropolysaccharide from the fruiting bodies of Morchella esculenta.

This study aimed to investigate the structural features, in vitro and in vivo antioxidant activities of a heteropolysaccharide from the fruiting bodies of Morchella esculenta (FMP-1). FMP-1 had an average molecular weight of 4.7 × 103 Da and consisted of mannose, glucose and galactose. By methylation and NMR analysis, the backbone of FMP-1 was deduced to be made up of 1,4-linked Glcp and 1,6-linked Galp. Hydroxyl, DPPH and superoxide radicals could be efficiently scavenged by FMP-1, with IC50 values of 74.26, 119.32 and 161.49 µg/mL, respectively. Furthermore, FMP-1 could significantly protect zebrafish embryos against AAPH-induced oxidative damage. Decrease in malformations and mortalities was observed along with the reduction of ROS production, NO production and cell death. The protective effects were by decreasing MDA content and increasing SOD, CAT and GSH-Px levels. The current work provided a good suggestion of the potential utilization of FMP-1 as an attractive natural antioxidant.

Structural elucidation of three antioxidative polysaccharides from Tricholoma lobayense.

Tricholoma lobayense is a nutritious mushroom with great health benefits. Three polysaccharides with purity higher than 99% were successfully extracted from Tricholoma lobayense. The molecular weights of TLH-1, TLH-2 and TLH-3 were determined to be 8.43×105，5.36×105 and 4.53×103Da, respectively. The backbones of TLH-1 and TLH-2 were mainly composed of 1,4-linked α-d-glucopyranosyl. However, polysaccharide TLH-3 was found to be a highly branched glucogalactan, which is made up of 1,3-linked α-d-glucopyranosyl branched at C-6 and 1,3-linked ß-d-galactopyranosyl. In vitro antioxidant activity assays revealed that TLH-3 exhibited highest antioxidant activities among the polysaccharides from Tricholoma lobayense, which were comparable to those of ascorbic acid. The results suggested that the outstanding antioxidant activities of TLH-3 might depend on its low molecular weight, high branch degree, versatile linkage types and complex conformation. These characteristics make TLH-3 an attractive natural antioxidant for food and pharmaceutical applications.

Synthesis, in silico and in vivo blood brain barrier permeability of ginkgolide B cinnamate.

Ginkgolide B, one of the important components of Ginkgo biloba extracts, has been revealed to exhibit great potential in therapy of cerebrovascular diseases. However the lack of permeability greatly limited it from further clinical application. Based on the prediction model for blood brain barrier (BBB) permeation, herein a potential brain-targeting analog ginkgolide B cinnamate (GBC) was successfully synthesized and characterized. After intravenous administration of GBC or GB, liquid chromatography tandem mass spectrometry (LC-MS/MS) was conducted to determine the analog in rat plasma and brain. The results showed that GBC had a significant increase in BBB permeability. A significant 1.61-times increase in half-life was observed for GBC and the drug targeting index (DTI) value was calculated to be 9.91. The experiment results matched well with the predicted one, which revealed that BBB permeability prediction model combined with in vivo study could be used as a quick, feasible and efficient tool for brain-targeting drug design.

Immunohistochemistry and Fluorescence In Situ Hybridization Can Inform the Differential Diagnosis of Low-Grade Noninvasive Urothelial Carcinoma with an Inverted Growth Pattern and Inverted Urothelial Papilloma.

Urothelial carcinoma (UC) comprises a heterogeneous group of epithelial neoplasms with diverse biological behaviors and variable clinical outcomes. Distinguishing UC histological subtypes has become increasingly important because prognoses and therapy can dramatically differ among subtypes. In clinical work, overlapping morphological findings between low-grade noninvasive UC (LGNUC), which exhibits an inverted growth pattern, and inverted urothelial papilloma (IUP) can make subclassification difficult. We propose a combination of immunohistochemistry (IHC) and molecular cytogenetics for subtyping these clinical entities. In our study, tissue microarray immunohistochemical profiles of Ki-67, p53, cytokeratin 20 (CK20) and cyclinD1 were assessed. Molecular genetic alterations such as the gain of chromosomes 3, 7 or 17 or the homozygous loss of 9p21 were also assessed for their usefulness in differentiating these conditions. Based on our analysis, Ki-67 and CK20 may be useful for the differential diagnosis of these two tumor types. Fluorescence in situ hybridization (FISH) can also provide important data in cases in which the malignant nature of an inverted urothelial neoplasm is unclear. LGNUC with an inverted growth pattern that is negative for both Ki-67 and CK20 can be positively detected using FISH.

Synthesis and transmembrane anion/cation symport activity of a rigid bis(choloyl) conjugate functionalized with guanidino groups.

A rigid bis(choloyl) conjugate functionalized with guanidino groups was synthesized and fully characterized on the basis of NMR ((1)H and (13)C) and ESI MS (LR and HR) data. Its transmembrane ionophoric activity across egg-yolk l-α-phosphatidylcholine-based liposomal membranes was investigated by means of chloride ion selective electrode technique and pH discharge assay. The data indicate that under the assay conditions, this conjugate was capable of promoting the transport of anions, presumably via a cation/anion symport process. A Hill analysis reveals that two molecules of this compound are assembled into the transport-active species.

Synthesis and potent ionophoric activity of a squaramide-linked bis(choloyl) conjugate.

A squaramide-linked bis(choloyl) conjugate was synthesized and fully characterized on the basis of NMR ((1)H and (13)C) and ESI MS (LR and HR) data. Fluorescence and chloride ion selective electrode assays indicate that this compound exhibits potent ionophoric activity across egg-yolk L-α-phosphatidylcholine-based liposomal membranes, presumably via an anion-modulating anion-cation co-transport/symport process. A Hill analysis reveals that three molecules of this compound are assembled into the transport-active species.

Synthesis and anionophoric activities of dimeric polyamine-sterol conjugates: the impact of rigid vs. flexible linkers.

Two dimeric spermine-choloyl conjugates were synthesized and found to be capable of promoting the transport of anions across egg-yolk L-α-phosphatidylcholine-based liposomal membranes, via an anion-exchange mechanism and with moderate selectivity with respect to monoanionic ions. A Hill analysis indicated that these two conjugates exhibited similar aggregation behaviors. However, the conjugate bearing a rigid p-bis(aminomethyl)benzene moiety functioned more efficiently than the analogue having a flexible putrescine linker.

Selective hydrogenation of nitroaromatics by ceria nanorods.

Ceria (CeO2) nanorods with well-defined surface planes can be synthesized and utilized for the hydrogenation of nitroaromatics. The CeO2 nanorods containing a {110} plane can efficiently and selectively catalyse the hydrogenation of nitroaromatics with N2H4 as a reducing agent, while nano-ceria with a {100} or {111} plane shows poor performance for the reaction.