Characterization of a Panel of Constitutive Promoters from Lactococcus cremoris for Fine-Tuning Gene Expression.

Lactococcus cremoris (homotypic synonym: Lactococcus lactis) is receiving increasing attention as a prominent vehicle for the delivery of live vaccines. This can hardly be achieved without developing tools for the genetic manipulation of L. cremoris, and the paucity of studies on L. cremoris endogenous promoters has attracted our attention. Here, we report the discovery and characterization of 29 candidate promoters identified from L. cremoris subsp. cremoris NZ9000 by RNA sequencing analysis. Furthermore, 18 possible constitutive promoters were obtained by RT-qPCR screening from these 29 candidate promoters. Then, these 18 promoters were cloned and characterized by a reporter gene, gusA, encoding ß-glucuronidase. Eventually, eight endogenous constitutive promoters of L. cremoris were obtained, which can be applied to genetic manipulation of lactic acid bacteria.

Three-Component Radical Cross-Coupling: Asymmetric Vicinal Sulfonyl-Esterification of Alkenes Involving Sulfur Dioxide.

A novel catalytic system for radical cross-coupling reactions based on copper and chiral Pyridyl-bis(imidazole) (PyBim) ligands is described. It overcomes the challenges of chemoselectivity and enantioselectivity, achieving a highly enantioselective vicinal sulfonyl-esterification reaction of alkenes involving sulfur dioxide. This strategy involves the use of earth-abundant metal catalyst, mild reaction conditions, a broad range of substrates (84 examples), high yields (up to 97% yield), and exceptional control over enantioselectivity. The reaction system is compatible with different types of radical precursors, including O-acylhydroxylamines, cycloketone oxime esters, aryldiazonium salts, and drug molecules. Chiral ligand PyBim is identified as particularly effective in achieving the desired high enantioselectivity. Mechanistic studies reveal that copper/PyBim system plays a vital role in C─O coupling, employing an outer-sphere model. In addition, the side arm effect of ligand is observed.

[Construction and stability analysis of finite element model for spinal canal reconstruction with miniplates fixation].

OBJECTIVE: To establish the finite element model of spinal canal reconstruction and internal fixation,analysis influence of spinal canal reconstruction and internal fixation on spinal stability,and verify the effectiveness and reliability of spinal canal reconstruction and internal fixation in spinal canal surgery. METHODS: A 30-year-old male healthy volunteer with a height of 172 cm and weight of 75 kg was selected and his lumbar CT data were collected to establish a finite element model of normal lumbar L3-L5,and the results were compared with in vitro solid results and published finite element analysis results to verify the validity of the model. They were divided into normal group,laminectomy group and spinal canal reconstruction group according to different treatment methods. Under the same boundary fixation and physiological load conditions,six kinds of activities were performed,including forward bending,backward extension,left bending,right bending,left rotation and right rotation,and the changes of range of motion (ROM) of L3-L4,L4-L5 segments and overall maximum ROM of L3-L5 were analyzed under the six conditions. RESULTS: The ROM displacement range of each segment of the constructed L3-L5 finite element model was consistent with the in vitro solid results and previous literature data,which confirms the validity of the model. In L3-L4,ROM of spinal canal reconstruction group was slightly increased than that of normal group during posterior extension(>5% difference),and ROM of other conditions was similar to that of normal group(<5% difference). ROM in laminectomy group was significantly increase than that in normal group and spinal canal reconstruction group under the condition of flexion,extension,left and right rotation. In L4-L5,ROM in spinal canal reconstruction group was similar to that in normal group(<5% difference),while ROM in laminectomy group was significantly higher than that in normal group and spinal canal reconstruction group(>5% difference). In the overall maximum ROM of L3-L5,spinal canal reconstruction group was only slightly higher than normal group under the condition of posterior extension(>5% difference),while laminectomy was significantly higher than normal group and spinal canal reconstruction group under the condition of anterior flexion,posterior extension,left and right rotation(>5% difference). The changes of each segment ROM and overall ROM of L3-L5 showed laminectomy group>spinal canal reconstruction group>normal group. CONCLUSION: Laminectomy could seriously affect biomechanical stability of the spine,but application of spinal canal reconstruction and internal fixation could effectively reduce ROM displacement of the responsible segment of spine and maintain its biomechanical stability.

Identification of bioactive compounds and inhibitory effects of TNF-α and COX-2 in the extract from cultured three-spot seahorse (H. trimaculatus).

Three-spot seahorse (Hippocampus trimaculatus) has been consumed as traditional Chinese medicine in Asian society. This study was designed to analyze the bioactive compounds of the solvent extracts from cultured three-spot seahorse by high pressure liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI/MS/MS). Subsequently, their biological activities were evaluated and confirmed by cell modes and Western blot analysis. Experimental results indicated that taurine and arginine were the primary bioactive compounds identified and quantified without pre- or post-column derivatization within 20 min retention time. The analytical method was established and validated with intraday/interday RSD from 0.25% to 3.34% and with recovery from 87.8% to 91.2%. As compared to other extracts, water layer extract (WLE) contained the most taurine and arginine contents of 6.807 and 0.437 mg/g (dry basis), respectively. In the meanwhile, WLE also showed anti-inflammatory activity on LPS-induced NO production and inhibited the protein expression of TNF-α and COX-2 by Western blot analysis with better cell viability.

Comparison on Ameliorating Effect of Arisaematis Rhizoma and Arisaematis Rhizoma Preparatum on Allergic Asthma in Rats / 中国实验方剂学杂志

ObjectiveTo investigate the therapeutic effects and difference in the effects of Arisaematis Rhizoma （AR） before and after processing （i.e.， Arisaematis Rhizoma Preparatum， ARP） with Zingiberis Rhizoma Recens-Alumen on allergic asthma in rats and to provide a basis for the theory of processing improving the efficacy. MethodA rat model of allergic asthma was established in 70 SD rats by intraperitoneal injection of ovalbumin （OVA）-aluminum hydroxide. The rats were administrated with the aqueous extracts of AR （1.2， 0.3 g∙kg-1） and ARP （1.2， 0.3 g∙kg-1） aqueous extracts by gavage， and montelukast sodium （0.001 g∙kg-1） was used as the positive drug. The T helper cell type 1/type 2 （Th1/Th2） ratio in the serum and bronchoalveolar lavage fluid （BALF） and percentages of inflammatory cells in BALF were determined. Polymerase chain reaction （PCR） was employed to determine the mRNA level of mucin 5AC （MUC5AC） in the lung tissue. The pathological changes in the lung tissue were observed by hematoxylin-eosin （HE） staining and PAS staining. Immunohistochemical assay was employed to measure the expression of c-Jun amino-terminal kinase （JNK）， extracellular signal regulated protein kinase （ERK）， and p38 mitogen-activated protein kinase （p38 MAPK） in rat lung tissue. Western blot was employed to determine the protein levels of ERK， p-ERK， JNK， p-JNK， p38， p-p38 in the lung tissue. The effects of AR and ARP were compared based on overall desirability. ResultCompared with the blank group， the levels of interleukin-12 （IL-12） and γ interferon （IFN-γ） in serum and BALF of rats in the model group were significantly lower （P<0.05， P<0.01）， and the levels of interleukin-4 （IL-4）， interleukin-5 （IL-5） and interleukin-13 （IL-13） were significantly higher （P<0.05， P<0.01）. Compared with the model group， the serum and BALF contents of IL-12 and IFN-γ in rats in the montelukast sodium group， high-dose AR group and high-dose ARP group were significantly higher （P<0.05， P<0.01）， and the contents of IL-4， IL-5 and IL-13 were significantly lower （P<0.05， P<0.01）， and the serum contents of IFN-γ in rats in the low-dose AR group and low-dose ARP group were in BALF was significantly higher （P<0.05） and IL-4 and IL-13 were significantly lower （P<0.05， P<0.01）， the percentages of macrophages， lymphocytes， neutrophils， and eosinophils were reduced in BALF， and the expression of JNK/ERK/p38 MAPK signaling pathway and MUC5AC protein was inhibited in lung tissues. Overall assessment of the normalized analysis revealed that the ARP group was slightly more potent than the AR group after administration of the same dose. ConclusionAR and ARP can effectively treat allergic asthma by inhibiting JNK/ERK/p38 MAPK signaling pathway， and the effect is better after concoction， which can provide data support for its "concoction efficiency".

Mechanochemical Synthesis of Aryl Fluorides by Using Ball Milling and a Piezoelectric Material as the Redox Catalyst.

Aryl fluorides are important structural motifs in many pharmaceuticals. Although the Balz-Schiemann reaction provides an entry to aryl fluorides from aryldiazonium tetrafluoroborates, it suffers from drawbacks such as long reaction time, high temperature, toxic solvent, toxic gas release, and low functional group tolerance. Here, we describe a general method for the synthesis of aryl fluorides from aryldiazonium tetrafluoroborates using a piezoelectric material as redox catalyst under ball milling conditions in the presence of Selectfluor. This approach effectively addresses the aforementioned limitations. Furthermore, the piezoelectric material can be recycled multiple times. Mechanistic investigations indicate that this fluorination reaction may proceed via a radical pathway, and Selectfluor plays a dual role as both a source of fluorine and a terminal reductant.

Copper-Catalyzed 1,4-Trifluoromethylthio-Arylsulfonylation of 1,3-Enynes via the Insertion of Sulfur Dioxide.

A copper-catalyzed trifluoromethylthio-arylsulfonylation between 1,3-enynes, AgSCF3, aryldiazonium tetrafluoroborates, and SO2 (from SOgen) is presented, which could introduce sulfone, SCF3, and allene moieties into one molecule simultaneously. This strategy features mild reaction conditions, good substrate compatibility, and excellent regioselectivity. The products obtained have the potential for further conversion into other valuable compounds. Initial investigations into the reaction mechanism suggest that it may proceed via a radical pathway. Notably, SOgen was proven as a uniquely effective SO2 surrogate in this transformation.

Association of family function and suicide risk in teenagers with a history of self-harm behaviors: mediating role of subjective wellbeing and depression.

Background: A history of self-harm behaviors is closely associated with subsequent suicide death. Although many factors associated with suicide have been identified, it remains unclear how these factors interact to influence suicide risk, especially among teenagers with a history of self-harm behaviors. Methods: Data were collected from 913 teenagers with a history of self-harm behaviors through a cross-sectional study. The Family Adaptation, Partnership, Growth, Affection, and Resolve index was used to assess teenagers' family function. The Patient Health Questionnaire-9 and the Generalized Anxiety Disorder-7 were used to evaluate depression and anxiety in teenagers and their parents, respectively. The Delighted Terrible Faces Scale was used to assess teenagers' perception of subjective wellbeing. The Suicidal Behaviors Questionnaire-Revised was used to evaluate teenagers' suicide risk. Student's t-test, one-way ANOVA, multivariate linear regression, Pearson's correlation, and a structural equation model (SEM) were applied to data analysis. Results: Overall, 78.6% of teenagers with a history of self-harm behaviors were at risk for possible suicide. Female gender, severity of teenagers' depression, family function, and subjective wellbeing were significantly associated with suicide risk. The results of SEM suggested that there was a significant chain mediation effect of subjective wellbeing and depression between family function and suicide risk. Conclusion: Family function was closely associated with suicide risk in teenagers with a history of self-harm behaviors, and depression and subjective wellbeing were sequential mediators in the association between family function and suicide risk.

A New Type of Endometrial Cancer Models in Mice Revealing the Functional Roles of Genetic Drivers and Exploring their Susceptibilities.

Endometrial cancer (EC) is the most common female reproductive tract cancer and its incidence has been continuously increasing in recent years. The underlying mechanisms of EC tumorigenesis remain unclear, and efficient target therapies are lacking, for both of which feasible endometrial cancer animal models are essential but currently limited. Here, an organoid and genome editing-based strategy to generate primary, orthotopic, and driver-defined ECs in mice is reported. These models faithfully recapitulate the molecular and pathohistological characteristics of human diseases. The authors names these models and similar models for other cancers as organoid-initiated precision cancer models (OPCMs). Importantly, this approach can conveniently introduce any driver mutation or a combination of driver mutations. Using these models,it is shown that the mutations in Pik3ca and Pik3r1 cooperate with Pten loss to promote endometrial adenocarcinoma in mice. In contrast, the Kras G12D mutati led to endometrial squamous cell carcinoma. Then, tumor organoids are derived from these mouse EC models and performed high-throughput drug screening and validation. The results reveal distinct vulnerabilities of ECs with different mutations. Taken together, this study develops a multiplexing approach to model EC in mice and demonstrates its value for understanding the pathology of and exploring the potential treatments for this malignancy.

Characterization of structural and functional properties of soy protein isolate and sodium alginate interpenetrating polymer network hydrogels.

BACKGROUND: This study used enzymatic and Ca2+ cross-linking methods to prepare edible soy protein isolate (SPI) and sodium alginate (SA) interpenetrating polymer network hydrogels to overcome the disadvantages of traditional interpenetrating polymer network (IPN) hydrogels, such as poor performance, high toxicity, and inedibility. The influence of changes in SPI and SA mass ratio on the performance of SPI-SA IPN hydrogels was investigated. RESULTS: Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) were used to characterize the structure of the hydrogels. Texture profile analysis (TPA), rheological properties, swelling rate, and Cell Counting Kit-8 (CCK-8) were used to evaluate physical and chemical properties and safety. The results showed that, compared with SPI hydrogel, IPN hydrogels had better gel properties and structural stability. As the mass ratio of SPI-SA IPN changed from 1:0.2 to 1:1, the gel network structure of hydrogels also tended to be dense and uniform. The water retention and mechanical properties of these hydrogels, such as storage modulus (G'), loss modulus (G"), and gel hardness increased significantly and were greater than those of the SPI hydrogel. Cytotoxicity tests were also performed. The biocompatibility of these hydrogels was good. CONCLUSIONS: This study proposes a new method to prepare food-grade IPN hydrogels with mechanical properties of SPI and SA, which may have strong potential for the development of new foods. © 2023 Society of Chemical Industry.

Structural basis of the substrate recognition and inhibition mechanism of Plasmodium falciparum nucleoside transporter PfENT1.

By lacking de novo purine biosynthesis enzymes, Plasmodium falciparum requires purine nucleoside uptake from host cells. The indispensable nucleoside transporter ENT1 of P. falciparum facilitates nucleoside uptake in the asexual blood stage. Specific inhibitors of PfENT1 prevent the proliferation of P. falciparum at submicromolar concentrations. However, the substrate recognition and inhibitory mechanism of PfENT1 are still elusive. Here, we report cryo-EM structures of PfENT1 in apo, inosine-bound, and inhibitor-bound states. Together with in vitro binding and uptake assays, we identify that inosine is the primary substrate of PfENT1 and that the inosine-binding site is located in the central cavity of PfENT1. The endofacial inhibitor GSK4 occupies the orthosteric site of PfENT1 and explores the allosteric site to block the conformational change of PfENT1. Furthermore, we propose a general "rocker switch" alternating access cycle for ENT transporters. Understanding the substrate recognition and inhibitory mechanisms of PfENT1 will greatly facilitate future efforts in the rational design of antimalarial drugs.

Enantioselective Intermolecular Radical Amidation and Amination of Benzylic C-H Bonds via Dual Copper and Photocatalysis.

A method for direct access to enantioenriched benzylic amides and carbamate-protected primary benzylamines by C-H functionalization is reported. The C-H substrate is used as limiting reagent with only a small excess of the unactivated amide or carbamate nucleophile. The enantioselective intermolecular dehydrogenative C-N bond formation is enabled by a combination of a chiral copper catalyst, a photocatalyst, and an oxidant, and it takes place under mild conditions, which allow for a broad substrate scope. The method is compatible with late-stage C-H functionalization, and it provides easy access to 15 N-labeled amides and amines starting from cheap 15 NH4 Cl.

Palladium-Catalyzed Carbonylative Hiyama-Denmark Reaction toward the Synthesis of Aryl Carbonyl-Containing Oxindoles.

A palladium-catalyzed domino Heck cyclization/carbonylative Hiyama-Denmark cross-coupling reaction between alkene-tethered aryl iodides and silylcarboxylic acids is presented. This reaction proceeds well without toxic carbon monoxide (CO) gas and has good functional group tolerance, providing an alternative access to carbonyl-containing oxindoles. In this transformation, silylcarboxylic acids play a dual role as a CO source and a nucleophile.

Metabonomics analysis on Terra Flavausta in treating diarrhea with spleen yang deficiency based on UPLC-Q-Exactive MS / 国际中医中药杂志

Objective:To study the effects of Terra Flavausta on diarrhea mice with spleen yang deficiency based on metabonomics. Methods:Totally 30 mice were divided into normal group, model group and Terra Flavausta group according to random number table method. Mice in the model group and Terra Flavausta group were treated by the method of "diet disorder + clearing fire with herbs bitter in flavour and cold in property" to establish the diarrhea model of spleen yang deficiency. After successful modeling, Terra Flavausta group received Zaoxintu Decoction 12.0 g/kg for gavage, while normal group and model group were given equal volume of distilled water for gavage, for consecutive 7 d. The serum metabolites of each mouse were analyzed and identified based on UPLC-Q-Exective-MS. The differential metabolites were characterized by principal component analysis and orthogonal partial least squares discriminant analysis, and the potential biomakers were screened, and the KEGG pathway enrichment analysis was performed. Results:Totally 110 different metabolites were screened under the positive and negative ion mode. Terra Flavausta can effectively reverse the disorder of serum metabolism in diarrhea mice with spleen yang deficiency, and has a significant callback effect on 12 potential biomarkers related to diarrhea with spleen yang deficiency. KEGG pathway enrichment mainly involved HIF-1 signaling pathway, ascorbate and aldarate metabolism, platelet activation, etc. Conclusion:Terra Flavausta may play the effect of warming spleen and relieving diarrhea through down-regulation of L-ascorbic acid affecting HIF-1 signal pathway, ascorbic acid and aldose metabolism pathway, vitamin digestion and absorption pathway, up-regulation of prostaglandins G2 and H2 affecting platelet activation pathway, and down-regulation of jasmonic acid α linolenic acid metabolic pathway.

Chemical Constituents of the Deep-Sea-Derived Penicillium citreonigrum MCCC 3A00169 and Their Antiproliferative Effects.

Six new citreoviridins (citreoviridins J-O, 1-6) and twenty-two known compounds (7-28) were isolated from the deep-sea-derived Penicillium citreonigrum MCCC 3A00169. The structures of the new compounds were determined by spectroscopic methods, including the HRESIMS, NMR, ECD calculations, and dimolybdenum tetraacetate-induced CD (ICD) experiments. Citreoviridins J-O (1-6) are diastereomers of 6,7-epoxycitreoviridin with different chiral centers at C-2-C-7. Pyrenocine A (7), terrein (14), and citreoviridin (20) significantly induced apoptosis for HeLa cells with IC50 values of 5.4 µM, 11.3 µM, and 0.7 µM, respectively. To be specific, pyrenocine A could induce S phase arrest, while terrein and citreoviridin could obviously induce G0-G1 phase arrest. Citreoviridin could inhibit mTOR activity in HeLa cells.

The dynamics of microbial community and flavor metabolites during the acetic acid fermentation of Hongqu aromatic vinegar.

In this study, we investigated the dynamics of microbial community and flavor metabolites during the traditional fermentation of Hongqu aromatic vinegar (HAV) and subsequently explored the potential relationship between microbiota and flavor metabolites. The microbiome analysis based on high-throughput sequencing (HTS) of amplicons demonstrated that Lactobacillus, Acetobacter and Clostridium were the dominant bacterial genera, while Alternaria, Candida, Aspergillus and Issatchenkia were the dominant fungal genera during the acetic acid fermentation (AAF) of HAV. A total of 101 volatile flavor compounds were identified through gas chromatography-mass spectrometry (GC-MS) during HAV fermentation, including esters (35), alcohols (17), aldehydes (11), acids (11), ketones (7), phenols (10), and others (10). Redundancy analysis (RDA) was used to reveal the correlation between microbiota and volatile flavor compounds. Lactobacillus and Acetobacter were the two bacterial genera that have the great influence on the production of volatile flavor components in HAV. Among them, Lactobacillus was positively correlated with a variety of ethyl esters, while Acetobacter positively contributed to the formation of several organic acids. Furthermore, the non-volatile metabolites were detected by ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). A total of 41 dipeptides were identified during HAV fermentation, and most of them may have sensory characteristics and biological activities. RDA showed that Aspergillus, Epicoccum, Issatchenkia, Candida and Malassezia were the most influential fungal genera on non-volatile metabolites. In particular, Epicoccum was first reported in Hongqu vinegar and showed a positive correlation with the production of various organic acids. In conclusion, this study provides a scientific basis for understanding the flavor generation mechanism of HAV, and may be valuable for developing effective techniques to select suitable strains to improve the flavor quality of HAV.

Iron-Catalyzed Sulfonylthiocyanation of α,ß-Unsaturated Amides/Esters via the Insertion of Sulfur Dioxide.

An iron-catalyzed four-component sulfonylthiocyanation between α,ß-unsaturated amides/esters, TMSNCS, aryldiazonium tetrafluoroborates, and sulfur dioxide (from SOgen) is demonstrated. This protocol is characterized by mild reaction conditions, good functional group compatibility, broad substrate scope, and good to excellent yields, providing a feasible method for the preparation of ß-thiocyanated sulfone compounds. The preliminary mechanism investigation shows that a radical pathway may be involved in the process.

Intestinal microbiomics and liver metabolomics insights into the preventive effects of chromium (III)-enriched yeast on hyperlipidemia and hyperglycemia induced by high-fat and high-fructose diet.

In recent years, organic chromium (III) supplements have received increasing attentions for their low toxicity, high bioavailability and wide range of health-promoting benefits. This study aimed to investigate the preventive effects of chromium (III)-enriched yeast (YCr) on high-fat and high-fructose diet (HFHFD)-induced hyperlipidemia and hyperglycemia in mice, and further clarify its mechanism of action from the perspective of intestinal microbiomics and liver metabolomics. The results indicated that oral administration of YCr remarkably inhibited the aberrant elevations of body weight, blood glucose and lipid levels, hepatic cholesterol (TC) and triglyceride (TG) levels caused by HFHFD. Liver histological examination showed that oral YCr intervention inhibited HFHFD induced liver lipid accumulation. Besides, 16S rDNA amplicon sequencing showed that YCr intervention was beneficial to ameliorating intestinal microbiota dysbiosis by altering the proportion of some intestinal microbial phylotypes. Correlation-based network analysis indicated that the key intestinal microbial phylotypes intervened by YCr were closely related to some biochemical parameters associated with glucose and lipid metabolism. Liver metabolomics analysis revealed that dietary YCr intervention significantly regulated the levels of some biomarkers involved in purine metabolism, glycerophospholipid metabolism, citrate cycle, pyrimidine metabolism, glycerophospholipid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and so on. Moreover, dietary YCr intervention regulated the mRNA levels of key genes associated with glucose, cholesterol, fatty acids and bile acids metabolism in liver. These findings suggest that dietary YCr intervention has beneficial effects on glucose and lipid metabolism by regulating intestinal microbiota and liver metabolic pathway, and thus can be served as a functional component to prevent hyperlipidemia and hyperglycemia.

Effects of the microbial community on the formation of volatile compounds and biogenic amines during the traditional brewing of Hongqu rice wine.

As a typical representative of Chinese rice wine (Huangjiu), Hongqu rice wine is famous for its red color, mellow taste and strong fragrance. However, due to the open brewing environment and traditional fermentation technology, there are some safety risks in traditional brewed Hongqu rice wine, such as a certain amount of biogenic amines. In this study, the dynamic changes and the differences of microbial communities and volatile flavor components between two types of Hongqu rice wine with high and low biogenic amine contents (LBAW and HBAW) during the traditional brewing were systematically investigated. The results showed that the total biogenic amine contents in LBAW and HBAW were 20.91 and 69.06 mg/L, respectively. The contents of putrescine, cadaverine, spermine and spermidine in HBAW were significantly higher than those in LBAW, and it was noteworthy that spermine content in HBAW was 17.62 mg/L, which was not detected in LBAW. In addition, the volatile flavor characteristics of the two kinds of Hongqu rice wine were obviously different. The contents of acetophenone, n-butyl butanoate and benzothiazole were obviously higher in HBAW, while the contents of isoamyl acetate, ethyl lactate, ethyl caprate and phenylethyl alcohol were significantly higher in LBAW. High-throughput sequencing of 16S/ITS amplicon revealed that Weissella, Kosakonia, Pantoea, Monascus, Saccharomyces and Millerozyma were the predominant microbial genera during the traditional brewing of HBAW, while Weissella, Kosakonia, Monascus, Saccharomyces and Issatchenkia were the predominant microbial genera during the traditional brewing of LBAW. Correlation analysis revealed that biogenic amines were significantly negatively correlated with unclassified_o_Saccharomycetales, Cyberlindnera, Zygoascus, Aspergillus and Acinetobacter, but positively correlated with Lactobacillus, Pediococcus, Millerozyma and Apiotrichum. In addition, we also found that Lactobacillus, Pediococcus and Saccharomyces were significantly positively correlated with most of the volatile flavor components, while Candida, Trichosporon and Monascus were significantly negatively correlated with most of the volatile flavor components. In addition, bioinformatical analysis based on PICRUSt demonstrated that the key enzymes for biogenic amine biosynthesis were more abundant in the microbial community of HBAW than LBAW. These findings demonstrate that the formations of volatile flavor and biogenic amines in Hongqu rice wine are influenced by microbial community during the fermentation. This work facilitates scientific understanding of the formation mechanism of biogenic amines, and may be useful to develop effective strategies to improve the quality of Hongqu rice wine.

Organic chromium derived from the chelation of Ganoderma lucidum polysaccharide and chromium (III) alleviates metabolic syndromes and intestinal microbiota dysbiosis induced by high-fat and high-fructose diet.

Organic chromium is of great interest and has become an important chromium supplement resource in recent years because of its low toxicity and easy absorption. In our previous study, we synthesized a novel organic chromium [GLP-Cr] through the chelation of Ganoderma lucidum polysaccharide and chromium (III). The purpose of this study was to investigate the beneficial effects of GLP-Cr on the improvement of metabolic syndromes (MetS) in mice fed with a high-fat and high-fructose diet (HFHFD) and its mechanism of action. The results indicated that oral administration of GLP-Cr inhibited the excessive exaltation of body weight, glucose tolerance, fasting blood glucose and lipid levels, hepatic total cholesterol (TC), triglyceride (TG) levels caused by HFHFD. Besides, 16S rRNA amplicon sequencing showed that GLP-Cr intervention evidently ameliorated intestinal microbiota dysbiosis by changing the proportions of some intestinal microbial phylotypes. In addition, correlation network-based analysis indicated that the key intestinal microbial phylotypes were closely related to biochemical parameters associated with MetS under GLP-Cr intervention. Liver metabolomics analysis suggested that GLP-Cr intervention significantly regulated the levels of some biomarkers involved in alpha-linolenic acid metabolism, fatty acid biosynthesis, steroid hormone biosynthesis, glycerophospholipid metabolism, glycerolipid metabolism, steroid hormone biosynthesis, primary bile acid biosynthesis, and so on. Moreover, GLP-Cr intervention regulated liver mRNA levels of key genes associated with glucose and lipid metabolism. The mRNA level of glucose transporter type 4 (Glut4) was markedly increased by GLP-Cr intervention, and the mRNA levels of phosphoenolpyruvate carboxykinase (Pepck) and glucose-6-phosphatase (G6Pase) in the liver were significantly decreased. Meanwhile, GLP-Cr intervention significantly decreased hepatic mRNA levels of cluster of differentiation 36 (Cd36), acetyl-CoA carboxylase 1 (Acc1) and sterol regulatory element binding protein-1c (Srebp-1c), indicating that GLP-Cr intervention inhibited the excessive accumulation of free fatty acids in the liver. These findings suggest that the prevention of hyperglycemia and dyslipidemia by GLP-Cr may be closely related to the regulation of gut microbial composition and hepatic metabolic pathways, thus GLP-Cr can be serving as a functional component in the prevention of MetS.