An inulin-type fructan CP-A from Codonopsis pilosula attenuates experimental colitis in mice by promoting autophagy-mediated inactivation of NLRP3 inflammasome.

Inulin-type fructan CP-A, a predominant polysaccharide in Codonopsis pilosula, demonstrates regulatory effects on immune activity and anti-inflammation. The efficacy of CP-A in treating ulcerative colitis (UC) is, however, not well-established. This study employed an in vitro lipopolysaccharide (LPS)-induced colonic epithelial cell model (NCM460) and an in vivo dextran sulfate sodium (DSS)-induced colitis mouse model to explore CP-A's protective effects against experimental colitis and its underlying mechanisms. We monitored the clinical symptoms in mice using various parameters: body weight, disease activity index (DAI), colon length, spleen weight, and histopathological scores. Additionally, molecular markers were assessed through enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence (IF), immunohistochemistry (IHC), and Western blotting assays. Results showed that CP-A significantly reduced reactive oxygen species (ROS), tumor necrosis factor-alpha (TNF-α), and interleukins (IL-6, IL-1ß, IL-18) in LPS-induced cells while increasing IL-4 and IL-10 levels and enhancing the expression of Claudin-1, ZO-1, and occludin proteins in NCM460 cells. Correspondingly, in vivo findings revealed that CP-A administration markedly improved DAI, reduced colon shortening, and decreased the production of myeloperoxidase (MPO), malondialdehyde (MDA), ROS, IL-1ß, IL-18, and NOD-like receptor protein 3 (NLRP3) inflammasome-associated genes/proteins in UC mice. CP-A treatment also elevated glutathione (GSH) and superoxide dismutase (SOD) levels, stimulated autophagy (LC3B, P62, Beclin-1, and ATG5), and reinforced Claudin-1 and ZO-1 expression, thereby aiding in intestinal epithelial barrier repair in colitis mice. Notably, the inhibition of autophagy via chloroquine (CQ) diminished CP-A's protective impact against colitis in vivo. These findings elucidate that CP-A's therapeutic effect on experimental colitis possibly involves mitigating intestinal inflammation through autophagy-mediated NLRP3 inflammasome inactivation. Consequently, inulin-type fructan CP-A emerges as a promising drug candidate for UC treatment.

Dietary L-carnitine supplementation changes lipid metabolism and glucose utilization of Rhynchocypris lagowskii fed diets with different lipid sources.

The widely available crop oil is an effective alternative to the increasingly scarce marine fish oil. However, simple alternative strategies have led to declining growth and the edible value of farmed fish. It is worthwhile to explore the effects of micro supplements in diets to improve the tolerance of fish to different dietary lipid sources, which finally optimizes the feeding strategies. This study aimed to investigate the regulation of L-carnitine and dietary oil conditions on nutrient composition, lipid metabolism, and glucose regulation of Rhynchocypris lagowskii. Four diets were prepared according to fish oil, fish oil supplemented with L-carnitine, corn oil, and corn oil supplemented with L-carnitine, and FO, LCFO, CO, and LCCO were labeled, respectively. R. lagowskii was fed experimental diets for 8 weeks, and the glucose tolerance test was performed. The CO diet significantly resulted in higher crude lipid content in muscle but a lower level of serum lipid parameters of R. lagowskii than the FO diet. However, dietary L-carnitine supplementation significantly reduced the crude lipid content in the hepatopancreas and muscle of the fish fed with the CO diet yet increased the serum lipid parameters. Additionally, the crude lipid content of muscle was reduced in the fish fed with an FO diet supplemented with L-carnitine. Compared with the FO diet, the CO diet significantly reduced the ratio of n3/n6 polyunsaturated fatty acid in the hepatopancreas and muscle of R.lagowskii. Dietary L-carnitine supplementation significantly reduced the contents of total saturated fatty acids and total monounsaturated fatty acids in hepatopancreas under both dietary lipid sources. The CO diet significantly up-regulated the expression of genes related to lipid uptake and adipogenesis in hepatopancreas, including lipoprotein lipase (lpl), acetyl-coenzyme A carboxylase alpha (accα), and sterol regulatory element binding protein-1 (srebp1), compared with the FO diet. While dietary L-carnitine supplementation significantly down-regulated the expressions of lpl, accα, srebp1, and fatty acid synthase in hepatopancreas and muscle of fish under both dietary lipid sources, along with up-regulated expression of carnitine palmitoyltransferase 1 in hepatopancreas. Moreover, the fish fed with a CO diet significantly increased the expression of glucose uptake and clearance and significantly down-regulated the expressions of glucose regulation-related genes, including glucose transporter 1, glycogen synthase 1, and phosphofructokinase in hepatopancreas and muscle, resulting in slower glucose uptake and clearance than fish fed with FO diet. Nevertheless, dietary L-carnitine supplementation up-regulated the expression of gluconeogenesis-related genes, including glucose-6-phosphatase and phosphoenolpyruvate carboxykinase in the hepatopancreas of R. lagowskii under both dietary lipid sources. In conclusion, a higher dietary n6 PUFA resulted in lipid deposition, decreased serum lipid parameters, and limited serum glucose utilization of R. lagowskii. While the regulatory effect of L-carnitine on lipid metabolism and glucose utilization of R. lagowskii varies with dietary lipid sources and tissues.

Study on the Potential Mechanism of Tonifying Kidney and Removing Dampness Formula in the Treatment of Postmenopausal Dyslipidemia Based on Network Pharmacology, Molecular Docking and Experimental Evidence.

Background: Management of menopausal dyslipidemia is the main measure to reduce the incidence of cardiovascular disease in postmenopausal women. Tonifying Kidney and Removing Dampness Formula (TKRDF) is a traditional Chinese medicine (TCM) formula that ameliorates dyslipidemia in postmenopausal women. This study applied network pharmacology, molecular docking, and in vitro and in vitro experiments to investigate the underlying mechanism of TKRDF against postmenopausal dyslipidemia. Methods: Network pharmacology research was first conducted, and the active compounds and targets of TKRDF, as well as the targets of postmenopausal dyslipidemia, were extracted from public databases. Protein-protein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to identify the potential targets and signaling pathways of TKRDF in postmenopausal dyslipidemia. Molecular docking was then performed to evaluate the combination of active compounds with principal targets. Finally, an ovariectomized rat model was used for the in vivo experiment and alpha mouse liver 12 (AML12) cells treated with palmitic acid were used for the in vitro experiments to provide further evidence for the research. Results: Based on network pharmacology analysis, we obtained 78 active compounds from TKRDF that acted on 222 targets of postmenopausal dyslipidemia. The analysis results indicated that IL6, TNF, VEGFA, AKT1, MAPK3, MAPK1, PPARG and PIK3CA, etc., were the potentially key targets, and the PI3K/AKT signaling pathway was the possibly crucial pathway for TKRDF to treat postmenopausal dyslipidemia. Molecular docking suggested that the active compounds have good binding activity with the core targets. The in vivo and in vitro experiments demonstrated that TKRDF ameliorates postmenopausal dyslipidemia by regulating hormone levels, inhibiting inflammation, promoting angiogenesis and inhibiting lipid synthesis, which appear to be related to TKRDF's regulation of the ERK1/2 and PI3K/AKT signaling pathways. Conclusion: This study clarified the active ingredients, potential targets, and molecular mechanisms of TKRDF for treating postmenopausal dyslipidemia. It also provided a feasible method to uncover the scientific basis and therapeutic mechanism for prescribing TCM in the treatment of diseases.

Early evidence for beer drinking in a 9000-year-old platform mound in southern China.

Alcoholic beverages played an essential role in rituals in ancient societies. Here we report the first evidence for beer drinking in the context of burial ritual in early Holocene southern China. Recent archaeological investigations at Qiaotou (9,000-8,700 cal. BP) have revealed a platform mound containing human burials and high concentrations of painted pottery, encircled by a human-made ditch. By applying microfossil (starch, phytolith, and fungi) residue analysis on the pottery vessels, we found that some of the pots held beer made of rice (Oryza sp.), Job's tears (Coix lacryma-jobi), and USOs. We also discovered the earliest evidence for using mold saccharification-fermentation starter in beer making, predating written records by 8,000 years. The beer at Qiaotou was likely served in rituals to commemorate the burial of the dead. Ritualized drinking probably played an integrative role in maintaining social relationships, paving the way for the rise of complex farming societies four millennia later.

The origins of specialized pottery and diverse alcohol fermentation techniques in Early Neolithic China.

In China, pottery containers first appeared about 20000 cal. BP, and became diverse in form during the Early Neolithic (9000-7000 cal. BP), signaling the emergence of functionally specialized vessels. China is also well-known for its early development of alcohol production. However, few studies have focused on the connections between the two technologies. Based on the analysis of residues (starch, phytolith, and fungus) adhering to pottery from two Early Neolithic sites in north China, here we demonstrate that three material changes occurring in the Early Neolithic signal innovation of specialized alcoholic making known in north China: (i) the spread of cereal domestication (millet and rice), (ii) the emergence of dedicated pottery types, particularly globular jars as liquid storage vessels, and (iii) the development of cereal-based alcohol production with at least two fermentation methods: the use of cereal malts and the use of moldy grain and herbs (qu and caoqu) as starters. The latter method was arguably a unique invention initiated in China, and our findings account for the earliest known examples of this technique. The major ingredients include broomcorn millet, Triticeae grasses, Job's tears, rice, beans, snake gourd root, ginger, possible yam and lily, and other plants, some probably with medicinal properties (e.g., ginger). Alcoholic beverages made with these methods were named li, jiu, and chang in ancient texts, first recorded in the Shang oracle-bone inscriptions (ca. 3200 cal. BP); our findings have revealed a much deeper history of these diverse fermentation technologies in China.

Slowing disease progression in the SOD1 mouse model of ALS by blocking neuregulin-induced microglial activation.

There are no effective treatments to slow disease progression in ALS. We previously reported that neuregulin (NRG) receptors are constitutively activated on microglia in the ventral horns in both ALS patients and SOD1 mice and in the corticospinal tracts of ALS patients, and that NRG receptor activation occurs prior to significant clinical disease onset in SOD1 mice. Here, we hypothesize that blocking NRG signaling on microglia would slow disease progression in SOD1 mice using a targeted NRG antagonist (HBD-S-H4). Recombinant HBD-S-H4 directly delivered into the central nervous system (CNS) through implanted intracerebroventricular cannulas showed no signs of toxicity and significantly inhibited NRG receptor activation on microglia resulting in reduced microglial activation and motor neuron loss. The treatment also resulted in a delay in disease onset and an increase in survival. The therapeutic effect was dose-dependent that varied as a function of genetic background in two different strains of SOD1 mice. As a complementary drug delivery approach, transgenic mice expressing HBD-S-H4 driven by an astrocytic promoter (GFAP) had slower disease progression in a dose dependent manner, based on the level of HBD-S-H4 expression. These studies provide mechanistic insights into how NRG signaling on microglia may lead to disease progression and demonstrate the utility of a humanized fusion protein that blocks NRG as a novel therapeutic for human ALS.

Revealing a 5,000-y-old beer recipe in China.

The pottery vessels from the Mijiaya site reveal, to our knowledge, the first direct evidence of in situ beer making in China, based on the analyses of starch, phytolith, and chemical residues. Our data reveal a surprising beer recipe in which broomcorn millet (Panicum miliaceum), barley (Hordeum vulgare), Job's tears (Coix lacryma-jobi), and tubers were fermented together. The results indicate that people in China established advanced beer-brewing technology by using specialized tools and creating favorable fermentation conditions around 5,000 y ago. Our findings imply that early beer making may have motivated the initial translocation of barley from the Western Eurasia into the Central Plain of China before the crop became a part of agricultural subsistence in the region 3,000 y later.

[Inhibitory effect of Typhonium gigantewm Engl. on in vitro cultured human keloid fibroblasts].

OBJECTIVE: To study the inhibitory effect of Typhonium gigantewm Engl. (AEoTGE) on the proliferation and apoptosis of KFB in vitro and to survey the death rate. METHODS: Samples of hypertrophic scars were collected and cultured. Only 4-8 passage cells were selected for experiment. Inverted microscope and transmission electron microscope were used to observe the morphogenesis and ultrastructure of KFB. The KFB cells were treated with AEoTGE in different concentrations(3. 125,6.250, 12.500, 25.000, 50. 000,100.000 g/L) for 24 hours. The effect of AEoTGE on the proliferation and the IC50 of KFB was observed with MTT assay and EdU. The effect of AEoTGE on apoptosis of KFB was detected by flow cytometry. RESULTS: It showed that AEoTGE could inhibit the proliferation of KFB in an concentration-dependent style within the range of 3. 125-100.000 g/L. The AEoTGE could obviously increase the apoptosis rate of the KFB compared with blank control group(P <0.05). The IC50 of AEoTGE was 35 g/L. FITC-Annexin V/PI showed that apoptosis rate of KFB in the AEoTGE group was (72. 07 +/- 0. 70)% , while it was 23. 5% in blank control group (P < 0. 05). CONCLUSIONS: AEoTGE could significantly inhibit the proliferating activity and induce apoptosis of KFB after co-culture for 24 hours. The IC50 is 35 g/L and the rate of apoptosis is (72.07 +/- 0.70)%.

Sodium desoxycholate-assisted capillary electrochromatography with methacrylate ester-based monolithic column on fast separation and determination of coumarin analogs in Angelica dahurica extract.

A rapid and sensitive CEC method with methacrylate ester-based monolithic column has been developed for separation and determination of five coumarins (byakangelicin, oxypeucedanin hydrate, xanthotoxol, 5-hydroxy-8-methoxypsoralen and bergapten) in Angelica dahurica extract. Surfactant sodium desoxycholate (SDC) was introduced into the mobile phase as the pseudostationary to dynamically increase the selectivity of analytes instead of increasing the hydrophobicity of stationary phase. In addition, other factors, pH of phosphate buffer, ACN content and applied voltage, for instance, have also an obvious effect on the resolution but little on the retention time. Satisfactory separation of these five coumarins was achieved within 6 min under a 30:70 v/v ACN-buffer containing 20 mM sodium dihydrogen phosphate (NaH(2) PO(4) ) and 0.25 mM SDC at pH 2.51. The RSDs of intraday and interday for relative peak areas were less than 3.0% and 4.7%, respectively; and the recoveries were between 87.5% and 95.0%. The LODs were lower than 0.15 µg/mL and the LOQs were lower than 0.30 µg/mL, respectively, while calibration curves showed a good linearity (r(2) > 0.9979). Finally, five target coumarins from the crude extracts of A. dahurica were separated, purified, and concentrated by D-101 macroporous resin, and were successfully separated and quantitatively determined within 6 min.

Fingerprint development for Ginkgo biloba extracts by pressurized capillary electrochromatography: comparison of column types.

A pressurized capillary electrochromatographic (pCEC) fingerprint of Ginkgo biloba leaf extract was developed on three different types of capillary columns. A commercial column packed with 3-microm particles and an in-house column packed with 5-microm particles were investigated for their performance. Additionally, a monolithic column was included in the fingerprint study as a potential alternative to the conventional packed columns. The effects of experimental parameters, such as the composition of the mobile phase, the concentration and pH of the buffer, and the applied voltage, were studied. Binary mobile phases consisting of acetonitrile and a 5 mM sodium dihydrogen phosphate electrolyte at pH 2.8 were used in gradient elution mode with an applied voltage of 5 kV. Under optimal gradient conditions, at least 45 peaks were observed within 60 min on the commercial packed column, whereas only about 20 peaks were separated on the methacrylate-based monolithic and the in-house packed columns. The commercial column thus clearly outperforms the two other. However, the properties of the monolithic stationary phase still might be adapted (i.e., by changing the polymerization-mixture composition, the porosity, and thus the selectivity of the phase might be changed), which could lead to an improved efficiency.

Fast separation and determination of coumarins in Fructus cnidii extracts by CEC using poly(butyl methacrylate-co-ethylene dimethacrylate-co-[2-(methacryloyloxy)ethyl] trimethylammonium chloride) monolithic columns.

A rapid CEC method with poly(butyl methacrylate-co-ethylene dimethacrylate-co-[2-(methacryloyloxy)ethyl] trimethylammonium chloride) monolithic column has been developed for separation and determination of four coumarins (isopimpinelline, bergapten, imperatorin, and osthole) in Fructus cnidii extracts. The effect of polymerization condition including the monomers ratio and the porogens ratio were studied. The mobile-phase composition, such as the composition of organic solvent, the concentration and pH of buffer, was also optimized. Under the same condition (50% ACN and 50% of a 10 mM sodium dihydrogen phosphate electrolyte at pH 4.95), in contrast to 25 min of analysis time in HPLC and 10 min of analysis time in pCEC, a fast separation of these analytes was achieved in less than 5 min in CEC. Method validation was developed in accordance with the analytical procedures. Intra- and interday precisions (RSD) for relative retention time and peak area were less than 1.69 and 4.63%, and LODs were lower than 0.5 microg/mL. Calibration curves of four compounds also showed good linearity (r(2)>0.995). The mean recoveries ranged between 93.91 and 98.65%. With this CEC system, the quality of F. cnidii extracts from various resources was evaluated by determining the contents of the four coumarins.

Separation and determination of coumarins in Fructus cnidii extracts by pressurized capillary electrochromatography using a packed column with a monolithic outlet frit.

The pressurized capillary electrochromatography (pCEC) was utilized for the separation and determination of coumarins in Fructus cnidii extracts from 12 different regions. After a thorough study of analytical parameters such as acetonitrile content of the mobile phase, the concentration and pH of the buffer, and the applied voltage, a methodology was proposed to separate and determine six coumarins of F. cnidii extracts in less than 15 min. The experiments were performed in an in-house packed column with a monolithic outlet frit under the optimal conditions: pH 4.0 ammonium acetate buffer at 10 mM containing 50% acetonitrile at -6kV applied voltage. The calibration curves were linear in the range of 10.0-100.0 microg/mL for bergapten, 20.0-200.0 microg/mL for imperatorin, 5.0-400.0 microg/mL for osthole, 10.0-100.0 microg/mL for 2'-acetylangelicin, 10.0-200.0 microg/mL for oroselone, and 10.0-200.0 microg/mL for O-acetylcolumbianetin. The correlation coefficients were between 0.9967 and 0.9995. With this pCEC system, fingerprints of F. cnidii extracts were preliminarily established to distinguish three types of coumarins by characteristic peaks, and the quality of various sources of raw materials was evaluated by determining the contents of six coumarins.

Development of a quality evaluation method for Fructus schisandrae by pressurized capillary electrochromatography.

A pressurized CEC (pCEC) method with postcolumn detection cell had been developed for quantifying the lignans from Fructus schisandrae extracts. The effects of different experimental conditions, such as the ACN content of the mobile phase, the concentration and pH of the buffer, the applied voltage, and the supplementary pressure were studied. Five lignans (schisandrin, gomisin A, schisantherin C, deoxyschizandrin, schisandrin B) were baseline separated using a mobile phase of ACN-phosphate buffer (pH 5.4; 5 mM) (40:60 v/v) under -4 kV applied voltage. The method showed the satisfactory retention time and peak area repeatability. The calibration curves were linear in the range 50.0-1000.0 microg/mL for schisandrin, 20.0-500.0 microg/mL for gomisin A, 10.0-200.0 microg/mL for schisantherin C, 20.0-500.0 microg/mL for deoxyschizandrin, and 20.0-500.0 microg/mL for schisandrin B. The correlation coefficients were between 0.9978 and 0.9989. With this pCEC system, fingerprints of F. schisandrae were preliminarily established to distinguish two members S. chinensis (Turcz.) Baill. and S. sphenanthera Rehd. Et Wils. of F. schisandrae by characteristic peaks, and evaluate the quality of various sources of raw materials by determining the contents of the five lignans.