Search

Chinese rice wine (Huangjiu), a typical representative of traditional fermented foods in China, is distinguished for its unique flavor. Microorganisms play the most important role in the formation of the unique flavor of Huangjiu. The complex flavor components in Huangjiu are closely related to the metabolism of various microorganisms during the traditional brewing. Therefore, understanding the influence of microbiota on the formation of flavor components in Huangjiu is conducive to the production of Huangjiu with high-quality flavor. In recent years, many scholars have conducted a large number of studies on the microbial community and flavor metabolites during the traditional brewing process of Huangjiu, with the help of high-throughput sequencing (HTS) and mass spectrometry-based metabolomics. This review mainly summarized the current study progress of the microbial diversity of fermentation starters (Qu) and brewing process of Huangjiu, and explored the potential relationship between the microbial community and the characteristic flavor formation of Huangjiu. Through the comparative analysis of literatures, we found that Huangjiu has strong regional characteristics due to the differences in raw materials, Qu and traditional brewing techniques. Dynamics of microbial composition during the traditional brewing of Huangjiu in different regions of China are quite different, making Huangjiu in each region have their own flavor characteristics. Therefore, revealing the relationship between brewing microorganisms and flavor characteristics of Huangjiu is helpful to the directional regulation of flavor quality. According to the current research status, it is proposed that in the future, emerging technologies such as multi-omics, artificial intelligence, and machine learning should be applied to further improve the accuracy of the analysis of microbial flora and flavor components in Huangjiu, so as to accurately reveal the relationship between the microbial community and the formation of characteristic flavor of Huangjiu. Afterwards, high-throughput screening of excellent functional strains from the traditional brewing system should be applied to the industrial production of Huangjiu, so as to improve the overall flavor quality and commercial value.

This study aimed to investigate the beneficial effects of the oral administration of Lactobacillus brevis FZU0713-fermented Laminaria japonica (FLJ) on lipid metabolism and intestinal microbiota in hyperlipidemic rats fed with a high-fat diet (HFD). The results demonstrated that the oral administration of FLJ significantly inhibited obesity and improved the serum and hepatic biochemical parameters in HFD-fed rats. Histopathological results also indicated that FLJ intervention could significantly reduce the accumulation of lipid droplets in the liver induced by HFD feeding. Furthermore, FLJ intervention up-regulated the fecal short-chain fatty acid (SCFA) levels (mainly acetate, propionate and isobutyrate) in HFD-fed rats. Intestinal microbiota profiling by 16S rRNA gene sequencing revealed that FLJ intervention increased the relative abundance of Akkermansia, Collinsella, Ruminococcaceae_UCG-013, Defluviitaleaceae_UCG-011, Intestinimonas, Actinomyces and Tyzzerella, but decreased the abundance of Flavonifractor, Collinsella, Sporosarcina and Lacticigenium. Based on Spearman's correlation, the fecal levels of TC, TG, acetic acid and butyric acid were positively correlated with the relative abundance of Akkermansia and Ruminococcaceae_NK4A214, but negatively correlated with the relative amount of Flavonifractor and Collinsella. The metabolic function of intestinal microbiota predicted by PICRUSt analysis of 16S rRNA gene sequences demonstrated that primary and secondary bile acid biosyntheses, fatty acid biosynthesis, taurine and hypotaurine metabolism, arachidonic acid metabolism, glycolysis/gluconeogenesis, etc. were significantly down-regulated after 8 weeks of FLJ intervention. Additionally, FLJ intervention significantly regulated the hepatic mRNA levels (including BSEP, CYP7A1, LDLR, HMGCR, CD36 and SREBP1-C) involved in lipid metabolism and bile acid homeostasis. In conclusion, these findings support the possibility that Laminaria japonica fermented with probiotic Lactobacillus has the potential to reduce the disturbance of lipid metabolism by regulating intestinal microflora and liver gene expression profiles, so it can be employed as a potential functional food to prevent hyperlipidemia.

Inflammatory bowel disease (IBD) is a chronic intestinal inflammation that is currently incurable. Increasing evidence indicates that supplementation with probiotics could improve the symptoms of IBD. It is scientifically significant to identify novel and valid strains for treating IBD. It has been reported that the probiotic Lactobacillus paracasei L9 (L9), which is identified from the gut of healthy centenarians, can modulate host immunity and plays an anti-allergic role. Here, we demonstrated that L9 alleviates the pathological phenotypes of experimental colitis by expanding the abundance of butyrate-producing bacteria. Oral administration of sodium butyrate in experimental colitis recapitulates the L9 anti-inflammatory phenotypes. Mechanistically, sodium butyrate ameliorated the inflammatory responses by inhibiting the IL-6/STAT3 signaling pathway in colitis. Overall, these findings demonstrated that L9 alleviates the DSS-induced colitis development by enhancing the abundance of butyrate-producing bacterial strains that produce butyrate to suppress the IL-6/STAT3 signaling pathway, providing new insight into a promising therapeutic target for the remission of IBD.

Background: Brown adipose tissue (BAT) dissipates chemical energy as heat and has the potential to be a protective strategy to prevent obesity. microRNAs (miRNAs) are emerging as important posttranscriptional factors affecting the thermogenic function of BAT. However, the regulatory mechanism underlying miRNA-mediated energy metabolism in BAT is not fully understood. Here, we explored the roles of miR-22 in BAT thermogenesis and energy metabolism. Methods: Using global and conditional knockout mice as in vivo models and primary brown adipocytes as an in vitro system, we investigated the function of miR-22 in BAT thermogenesis in vivo and in vitro. Results: miR-22 expression was upregulated in BAT in response to cold exposure and during brown preadipocyte differentiation. Both global and conditional knockout mice displayed BAT whitening, impaired cold tolerance, and decreased BAT thermogenesis. Moreover, we found that miR-22 deficiency impaired BAT glycolytic capacity, which is critical for thermogenesis. The mechanistic results revealed that miR-22 activated the mTORC1 signaling pathway by directly suppressing Tsc1 and concomitantly directly suppressing Hif1an, an inhibitor of Hif1α, which promotes glycolysis and maintains thermogenesis. Conclusions: Our findings identify miR-22 as a critical regulator in the control of thermogenesis in BAT and as a potential therapeutic target for human metabolic disorders.

The present study investigated the composition of water extract of Grifola frondosa (GFWE) and its regulatory effects on glycolipid metabolism and gut microbiota in rats. Structural characteristics of GFWE were detected by high performance liquid chromatography and ultra-high-performance liquid chromatography with mass spectrometry. The hypoglycemic and hypolipidemic activities of GFWE were obtained by inhibition of weight gain, recovery of serum glucose and lipid, and reverse of liver tissue damage. GFWE has up-regulated the relative mRNA and protein expression levels of adenine monophosphate activated protein kinase-α, peroxisome proliferator-activated receptors-α, and glucokinase, while down-regulated sterol regulatory element-binding transcription factor-1c and acetyl CoA carboxylase. Moreover, GFWE increased the abundances of caecal bacteria Oscillibacter and Barnesiella, which showed a significant negative correlation with serum glucose and lipid parameters. And GFWE enhanced the levels of total bile acids and short-chain fatty acids in the cecum. These results indicated that GFWE could be provided as a potential candidate to regulate glycolipid metabolism and gut microbiota.

Mammary and extramammary Paget's Diseases (PD) are a malignant skin cancer characterized by the appearance of Paget cells. Although easily diagnosed, its pathogenesis remains unknown. Here, single-cell RNA-sequencing identified distinct cellular states, novel biomarkers, and signaling pathways - including mTOR, associated with extramammary PD. Interestingly, we identified MSI1 ectopic overexpression in basal epithelial cells of human PD skin, and show that Msi1 overexpression in the epidermal basal layer of mice phenocopies human PD at histopathological, single-cell and molecular levels. Using this mouse model, we identified novel biomarkers of Paget-like cells that translated to human Paget cells. Furthermore, single-cell trajectory, RNA velocity and lineage-tracing analyses revealed a putative keratinocyte-to-Paget-like cell conversion, supporting the in situ transformation theory of disease pathogenesis. Mechanistically, the Msi1-mTOR pathway drives keratinocyte-Paget-like cell conversion, and suppression of mTOR signaling with Rapamycin significantly rescued the Paget-like phenotype in Msi1-overexpressing transgenic mice. Topical Rapamycin treatment improved extramammary PD-associated symptoms in humans, suggesting mTOR inhibition as a novel therapeutic treatment in PD.

This study aimed to investigate the protective mechanism of common buckwheat (Fagopyrum esculentum Moench.) against nonalcoholic fatty liver disease (NAFLD) associated with dyslipidemia in mice that were fed a high-fat and high-cholesterol diet (HFD). Results showed that oral supplementation of common buckwheat significantly improved physiological indexes and biochemical parameters related to dyslipidemia and NAFLD in mice fed with HFD. Furthermore, the HFD-induced reductions in fecal short-chain fatty acids were reversed by common buckwheat intervention, which also increased the fecal bile acid (BA) abundance compared with HFD-induced hyperlipidemic mice. Liver metabolomics based on ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry demonstrated that common buckwheat supplementation made significant regulatory effects on the pentose phosphate pathway, starch and sucrose metabolism, primary BA biosynthesis, and so forth. The results of high-throughput sequencing revealed that common buckwheat supplementation significantly altered the structure of the intestinal microbiota in mice fed with HFD. The correlations between lipid metabolic parameters and intestinal microbial phylotypes were also revealed by the heatmap and network. Additionally, common buckwheat intervention regulated the mRNA expressions of genes responsible for liver lipid metabolism and BA homeostasis, thus promoting BA synthesis and excretion. These findings confirmed that common buckwheat has the outstanding ability of improving lipid metabolism and could be used as a potential functional food for the prevention of NAFLD and hyperlipidemia.

Ganoderic acid A (GA) is one of the most abundant triterpenoids in Ganoderma lucidum, and has been proved to possess a wide range of beneficial health effects. The aim of the current study is to investigate the amelioration effects and mechanism of GA on improving hyperlipidemia in mice fed a high-fat diet (HFD). The results showed that GA intervention significantly inhibited the abnormal growth of body weight and epididymal white adipose tissue (eWAT), prevented the hypertrophy of epididymal adipocytes, and ameliorated the biochemical parameters of serum and liver related to lipid metabolism in HFD-fed mice. Histological analysis also showed that the excessive accumulation of lipid droplets in the liver induced by HFD-feeding was greatly alleviated by GA intervention. In addition, GA intervention also increased the level of short chain fatty acids (SCFAs) in the intestine and promoted the excretion of bile acids (BAs) through feces. High-throughput sequencing of bacterial full-length 16S rDNA revealed that daily supplementation with GA made significant structural changes in the gut microbial population of mice fed with HFD, in particular modulating the relative abundance of some function related microbial phylotypes. The relationships between lipid metabolic parameters and gut microbial phylotypes were also revealed by correlation analysis based on a heatmap and network. The result showed that 46 key gut microbial phylotypes (OTUs) were markedly correlated with at least one lipid metabolic parameter. Moreover, UPLC-QTOF/MS-based liver metabolomics showed that 111 biomarkers (47 up-regulated metabolites and 64 down-regulated metabolites) were significantly changed after high-dose GA intervention (75 mg kg-1 day-1), compared with the HFD-fed hyperlipidemic mice. Metabolic pathway enrichment analysis of the differential hepatic metabolites demonstrated that GA intervention had significant regulatory effects on primary bile acid biosynthesis, fatty acid biosynthesis, amino sugar and nucleotide sugar metabolism, inositol phosphate metabolism, and so on. In addition, GA intervention regulated the mRNA levels of hepatic genes involved in fatty acid metabolism and bile acid homeostasis. These findings present new evidence supporting that GA from G. lucidum has the potential to alleviate lipid metabolic disorders and ameliorate the imbalance of gut microflora in a positive way.

Lactobacillus plantarum FZU3013, a probiotic previously isolated from the traditional brewing process of Hongqu rice wine, may have the beneficial effect of improving the disorders of lipid metabolism. This study aimed to investigate the role of L. plantarum FZU3013 in improving non-alcoholic fatty liver (NAFL) associated with hyperlipidemia in mice fed a high-fat diet. The results indicated that L. plantarum FZU3013 intervention significantly reduced the HFD-induced body weight gain and the abnormal levels of serum total triglycerides (TG), total cholesterol (TC) and low-density lipoprotein (LDL-C), and inhibited the excessive accumulation of liver lipids. In addition, L. plantarum FZU3013 also promoted the excretion of bile acids through feces. Metagenomic and multivariate statistical analysis revealed that L. plantarum FZU3013 made significant structural changes in the intestinal microbiome of the mice fed with HFD, in particular by modulating the relative abundance of some function related microbial phylotypes. Furthermore, ultra-performance liquid chromatography with quadruple-time of flight mass spectrometry (UPLC-QTOF/MS)-based liver metabolomics demonstrated that L. plantarum FZU3013 had a significant regulatory effect on the composition of liver metabolites in hyperlipidemic mice, especially on the levels of some important biomarkers involved in the pathways of glycerophospholipid metabolism, fatty acid degradation, fatty acid elongation, glycerolipid metabolism, primary bile acid biosynthesis, arachidonic acid metabolism, etc. Moreover, L. plantarum FZU3013 regulated the mRNA expression levels of the genes responsible for liver lipid and cholesterol metabolism. L. plantarum FZU3013 intervention increased the hepatic mRNA levels of cholesterol 7α-hydroxylase (CYP7A1) and the bile salt export pump (BSEP), suggesting enhanced bile acid synthesis and excretion from the liver. These findings present new evidence supporting that L. plantarum FZU3013 has the potential to improve lipid metabolism disorders through modulating specific intestinal microbial phylotypes and regulating hepatic lipid metabolism related genes, therefore it could be used as a potential functional food for the prevention of NAFL and hyperlipidemia.

Monascus spp. and its secondary metabolites have been widely applied in foods and medicines for thousands of years in eastern Asia. Nitrogen sources are essential nutrients for the growth and metabolism of Monascus spp. Our previous study found that inorganic nitrogen sources (especially NH4Cl and NH4NO3) promoted the biosynthesis of Monascus pigments (MPs) and inhibited the production of citrinin. The objective of the present study was to investigate the regulatory mechanism of inorganic nitrogen on the biosynthesis of MPs and citrinin by the comparative transcriptional approach (RNA sequencing combined with RT-qPCR). Results indicated that the submerged fermentation of M. purpureus M3103 with NH4Cl or NH4NO3 as the sole nitrogen source can significantly increase the yields of MPs (especially for Monascus orange and red pigments) and decrease citrinin production, compared with the organic nitrogen source (peptone group). Comparative transcriptomic profiling by RNA sequencing found that the numbers of differentially expressed genes (DEGs) between different experimental groups—M group (peptone group) vs. ML group (NH4Cl group), and M group (peptone group) vs. MX group (NH4NO3 group), were 722 and 1287, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that genes involved in carbon and nitrogen metabolism, biosynthesis of amino acids were up-regulated by NH4Cl and NH4NO3, which would produce more biosynthetic precursors for MPs. Whereas, the inorganic nitrogen source (both of NH4Cl and NH4NO3) down-regulated the expression levels of genes involved in tyrosine metabolism. In addition, NR analysis indicated that the essential genes and transcription factors involved in the biosynthesis pathway of citrinin were down-regulated by NH4Cl and NH4NO3. These results indicated that NH4Cl or NH4NO3 as a nitrogen source for M. purpureus M3103 can significantly promote the precursor synthesis of Monascus pigments, but reduce the transcription of polyketide synthase for citrinin, and therefore significantly increase Monascus pigments production and decrease citrinin formation. These findings will facilitate a comprehensive understanding of the regulatory mechanisms of inorganic nitrogen in the biosynthesis of secondary metabolites in M. purpureus, and would benefit the application of M. purpureus in the production of MPs.

Myoepithelial and luminal cells synergistically expand in the mammary gland during pregnancy, and this process is precisely governed by hormone-related signaling pathways. The bone morphogenetic protein (BMP) signaling pathway is now known to play crucial roles in all organ systems. However, the functions of BMP signaling in the mammary gland remain unclear. Here, we found that BMPR1a is upregulated by hormone-induced Sp1 at pregnancy. Using a doxycycline (Dox)-inducible BMPR1a conditional knockout mouse model, we demonstrated that loss of BMPR1a in myoepithelium results in compromised myoepithelial integrity, reduced mammary stem cells and precocious alveolar differentiation during pregnancy. Mechanistically, BMPR1a regulates the expression of p63 and Slug, two key regulators of myoepithelial maintenance, through pSmad1/5-Smad4 complexes, and consequently activate P-cadherin during pregnancy. Furthermore, we observed that loss of BMPR1a in myoepithelium results in the upregulation of a secreted protein Spp1 that could account for the precocious alveolar differentiation in luminal layer, suggesting a defective basal-to-luminal paracrine signaling mechanism. Collectively, these findings identify a novel role of BMP signaling in maintaining the identity of myoepithelial cells and suppressing precocious alveolar formation.

Shewanella and Pseudomonas are considered to be the main spoilage bacteria for chilled fish. This work focused on the spoilage and adhesion ability of two spoilage bacteria (Shewanella spp. S-1 and Pseudomonas spp. P-1), and the adhesion inhibitory effects of Bacillus subtilis against the spoilage bacteria in large yellow croaker (Pseudosciaena crocea). We aimed to develop a feasible solution to prolong the shelf life of large yellow croaker. The spoilage and adhesion characteristics of Shewanella spp. and Pseudomonas spp. strains were compared. It was found that Shewanella spp. induced more significant spoilage of large yellow croaker than Pseudomonas spp. at low temperature in vitro. In addition, Shewanella spp. demonstrated stronger adhesion to the intestinal mucus than Pseudomonas spp. in vitro. The results of a one-week feeding experiment showed that Shewanella was more likely to adhere to large yellow croaker intestines than Pseudomonas. The supplementation of B. subtilis strain BS08 effectively inhibited the proliferation of Shewanella and slowed the spoilage process of large yellow croaker.

Water activity (aw) and temperature are two pivotal environmental factors affecting Aspergillus flavus growth and aflatoxin production. Here, we found that AFB1 production on polished rice can occur over a wider range of temperature × aw levels than that on paddies. For fungal growth on polished rice, the optimum conditions were aw 0.92–0.96 and 28–37 °C. The maximum amounts of AFB1 on polished rice was observed at 33 °C and aw 0.96. Compared to 33 °C, all tested genes of A. flavus on polished rice were significantly up-regulated at 25 °C under aw 0.96. The late structural genes of pathway were significantly down-regulated at 37 °C under aw 0.96, although aflR and aflS and most of early structural genes were up-regulated. Compared to aw 0.96, most of pathway genes were significantly down-regulated at aw 0.90 and 0.99 under 33 °C, although two regulatory genes were up-regulated at aw 0.90.

This study aimed to investigate the effect of Spirulina mixed active substance (SPMX) on lipid metabolism and intestinal microbes in high-fat diet (HFD) rats. The effects of SPMX on body weight, serum lipids, pathological organization, gut microbiota and metabolites were analyzed. Results showed that body weight, serum lipids were significantly lower in SPMX group than that of HFD group, except HDL-C level. SPMX can enhance the restoration of fatty liver in HFD rats to a certain extent. SPMX obviously decreased the relative abundance of potential harmful bacteria. The mRNA expression levels of HMG-CoA, SREBP-1c and ACC in the liver tissue of the SPMX group were reduced remarkably compared with the HFD group. Spearman’s correlation analysis suggested that changes in feces microbiota was correlated with biochemical indicator. These findings suggested that SPMX could ameliorate lipid metabolism disorder, regulate intestinal microbial structure and can be used as lipid-lowering functional food.

Hong Qu glutinous rice wine (HQGRW), as one of the most typical representatives of Chinese rice wine, is generally brewed from glutinous rice by adding two traditional wine fermentation starters-Hong Qu (HQ) and Bai Qu (BQ). The objective of this study was to determine the microbial communities and volatile metabolites of different traditional fermentation starters for HQGRW, and elucidate the potential correlation between microbiota and volatile metabolites. Both heatmap and principal component analysis (PCA) revealed the significant variances in volatile profiles among different wine starters. Microbiological analysis based on high-throughput sequencing (HTS) technology demonstrated that both of bacterial and fungal communities varied significantly in different starters. HQ was dominated mainly by bacteria of Bacillus ginsengihumi (20.17%), Pantoea sp. (10.39%), Elizabethkingia sp. (5.52%), Streptococcus sp. (5.03%) Brevundimonas sp. (3.03%), Rickettsia prowazekii (2.94%), Thermus thermophilus (2.54%), Bacillus amyloliquefaciens (1.48%), Bacillus aryabhattai (1.42%); fungi of Monascus purpureus (39.7%), Aspergillus niger (27.35%), Xeromyces bisporus (8.39%), Aspergillus penicillioides (6.89%), Aspergillus flavus (2.33%) and Pichia farinose (0.79%). By contrast, BQ contained much higher proportions of bacteria of Lactococcus lactis (10.45%), Lactobacillus brevis (9.99%), Pediococcus pentosaceus (8.29%), Weissella paramesenteroides (6.69%), Lactobacillus fermentum (4.83%), Gluconobacter thailandicus (3.93%), Lactobacillus alimentarius (3.59%), fungi of Rhizopus arrhizus (31.47%), Saccharomycopsis fibuligera (27.86%), Aspergillus niger (20.81%), Issatchenkia orientalis (3.79%), Saccharomycopsis malanga (3.15%), Clavispora lusitaniae (2.29%), Candida tropicalis (1.47%), Saccharomyces cerevisiae (1.11%) and Rhizopus microsporus (0.57%). Furthermore, core functional microbiota that might contribute to volatile flavour development was explored through Spearman's correlation-based network analysis. Lactobacillus brevis, Lactobacillus alimentarius, Lactobacillus plantarum and Aspergillus niger were found to be strongly associated with acid compounds (FDR adjusted P 0.01), while Pichia sp., Candida sp., Monascus purpureus, Lactobacillus brevis and Lactobacillus alimentarius were positively correlated with concentrations of aromatic esters associated with fruity and floral notes (FDR adjusted P 0.01), implying that these microorganisms might make significant contributions to the flavour of rice wine. These findings demonstrated that the aromatic quality of HQGRW may be critically influenced by the microbiota in traditional fermentation starters. To conclude, this study would contribute to the development of novel defined starter cultures for improving the aromatic quality of HQGRW.

This study aimed to investigate the effects of polysaccharide peptides from Ganoderma lucidum (GLPP) on hyperlipidaemia and gut microbiota dysbiosis in high-fat diet (HFD)-exacerbated hypercholesterolemic rats. Results showed that oral administrations of GLPP markedly alleviated the dyslipidaemia through decreasing the serum levels of triglyceride (TG), cholesterol (TC), free fatty acids (FFA) and low-density lipoprotein cholesterol (LDL-C), and significantly suppressing hepatic lipid accumulation and steatosis. Metagenomic analysis revealed that GLPP supplementation produced significant structure changes on the intestinal microbiota in HFD-fed rats, in particular modulating the relative abundance of functionally relevant microbial phylotypes compared with the HFD group. The Spearman's correlation analysis revealed that the serum and hepatic lipid profiles were negatively correlated with Jeotgalicoccus, Ignavigranum, Sporosarcina, Bacteroides, Anaerovorax, Parasutterella, Alistipes and Alloprevotella, but positively correlated with Allobaculum, Phascolarctobacterium, Psychrobacter, Enterorhabdus, Blautia and Roseburia. Meanwhile, the GLPP treatment regulated the mRNA expression responsible for hepatic lipid metabolism and promoted fecal excretion of total bile acids (BAs). These findings indicated that GLPP ameliorate lipid metabolic disorders through modulating gut microbiota structure and regulating the genes involved in hepatic lipid and cholesterol metabolism.

The purpose of this study was to assess the potential effects of polysaccharides from edible mushroom Grifola frondosa (GFP) on lipid metabolic disorders and gut microbiota dysbiosis, and elucidate their possible regulatory mechanisms on lipid and cholesterol metabolism in high-fat diet (HFD)-exacerbated hyperlipidemic and hypercholesterolemic rats. Results showed that oral administration of GFP markedly alleviated dyslipidaemia through decreasing the serum levels of total triglycerides, total cholesterol, and free fatty acids, and significantly suppressing hepatic lipid accumulation and steatosis. Besides, the excretion of fecal bile acids was also promoted by oral administration of GFP. Metagenomic analysis revealed that GFP supplementation (400 mg kg-1 day-1) resulted in significant structure changes on gut microbiota in HFD-fed rats, in particular modulating the relative abundance of functionally relevant microbial phylotypes compared with the HFD group. Key microbial phylotypes responding to GFP intervention were identified to strongly correlate with the lipid metabolism disorder associated parameters using the correlation network based on Spearman's correlation coefficient. Serum and hepatic lipid profiles were found positively correlated with Clostridium-XVIII, Butyricicoccus and Turicibacter, but negatively correlated with Helicobater, Intestinimonas, Barnesiella, Parasutterella, Ruminococcus and Flavonifracter. Moreover, GFP treatment (400 mg kg-1 day-1) regulated the mRNA expression levels of the genes responsible for hepatic lipid and cholesterol metabolism. Oral supplementation of GFP markedly increased the mRNA expression of cholesterol 7α-hydroxylase (CYP7A1) and bile salt export pump (BSEP), suggesting an enhancement of bile acid (BA) synthesis and excretion from the liver. These findings illustrated that GFP could ameliorate lipid metabolic disorders through modulating specific gut microbial phylotypes and regulating hepatic lipid and cholesterol metabolism related genes, and therefore could be used as a potential functional food ingredient for the prevention or treatment of hyperlipidemia.

Diet can impact gut health and disease by modulating intestinal stem cells (ISCs). However, it is largely unknown how ISC niche responds to diet and influences ISC function. Here, we demonstrated that Lepr+ mesenchymal cells (MCs) surround intestinal crypts where ISCs and transit-amplifying (TA) cells localize. The abundance of these cells increased upon administration of a high-fat diet (HFD) but dramatically decreased upon fasting. Depletion of Lepr+ MCs resulted in fewer ISCs, compromised architecture of crypt-villus axis and impaired intestinal regeneration. Furthermore, IGF1 derived from Lepr+ MCs was found to be an important effector that promotes proliferation of ISCs. Overall, Lepr+ MCs sense diet alteration and function as a novel niche for ISCs via the stromal Igf1 - epithelial Igf1r axis. These findings revealed that Lepr+ MCs are an important mediator that links systemic diet changes to local ISC function and might provide a novel therapeutic target for gut diseases.

The aim of present study was to investigate the regulatory effects of an extract of the Grifola frondosa on glycolipid metabolism and the intestinal flora in high-fat diet fed rats. The ethyl acetate extract of G. frondosa (EAE), enriched with pseudobaptigenin and cyanidin 3-O-xylosylrutinoside, exerted a stronger inhibitory effect on lipase than the other polar fractions of G. frondosa in vitro. Supplementation with EAE (50 mg/kg) to rats for 8 weeks clearly ameliorated dyslipidaemia by suppressing the serum levels of glucose (Glu), total triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C), and hepatic fat accumulation. Furthermore, EAE supplementation normalized the intestinal flora structure in high-fat diet fed rats. The relevance analysis suggested strong associations of Anaerotruncus, Barnesiella, Clostridiaceae, Erysipelotrichaceae, Paraprevotella, Clostridium_XVIII, Coprococcus and Turicibacter with the biochemical indexes. Simultaneously, the EAE treatment intervened the expression levels of the genes associated with glycolipid metabolism. Our results indicated that pseudobaptigenin and cyanidin-3-O-xylosylrutinoside can be used as pharmaceutical agents for the prevention of the disorder of glycolipid metabolism.

MicroRNA-mediated post-transcriptional regulation plays key roles in stem cell self-renewal and tumorigenesis. However, the in vivo functions of specific microRNAs in controlling mammary stem cell (MaSC) activity and breast cancer formation remain poorly understood. Here we show that miR-31 is highly expressed in MaSC-enriched mammary basal cell population and in mammary tumors, and is regulated by NF-κB signaling. We demonstrate that miR-31 promotes mammary epithelial proliferation and MaSC expansion at the expense of differentiation in vivo. Loss of miR-31 compromises mammary tumor growth, reduces the number of cancer stem cells, as well as decreases tumor-initiating ability and metastasis to the lung, supporting its pro-oncogenic function. MiR-31 modulates multiple signaling pathways, including Prlr/Stat5, TGFβ and Wnt/β-catenin. Particularly, it activates Wnt/β-catenin signaling by directly targeting Wnt antagonists, including Dkk1. Importantly, Dkk1 overexpression partially rescues miR31-induced mammary defects. Together, these findings identify miR-31 as the key regulator of MaSC activity and breast tumorigenesis.

Mitotic inheritance of the DNA methylome is a challenging task for the maintenance of cell identity. Whether DNA methylation pattern in different genomic contexts can all be faithfully maintained is an open question. A replication-coupled DNA methylation maintenance model was proposed decades ago, but some observations suggest that a replication-uncoupled maintenance mechanism exists. However, the capacity and the underlying molecular events of replication-uncoupled maintenance are unclear. By measuring maintenance kinetics at the single-molecule level and assessing mutant cells with perturbation of various mechanisms, we found that the kinetics of replication-coupled maintenance are governed by the UHRF1–Ligase 1 and PCNA–DNMT1 interactions, whereas nucleosome occupancy and the interaction between UHRF1 and methylated H3K9 specifically regulate replication-uncoupled maintenance. Surprisingly, replication-uncoupled maintenance is sufficiently robust to largely restore the methylome when replication-coupled maintenance is severely impaired. However, solo-WCGW sites and other CpG sites displaying aging- and cancer-associated hypomethylation exhibit low maintenance efficiency, suggesting that although quite robust, mitotic inheritance of methylation is imperfect and that this imperfection may contribute to selective hypomethylation during aging and tumorigenesis.

The objective of this study was to investigate the effects of simvastatin (SIM) on lipid metabolism disorders and gut microbiota in high-fat diet-induced hyperlipidemic rats. The obtained results revealed that feeding rats with SIM (20 mg/kg/day) significantly decreased serum lipid level and inhibited hepatic lipid accumulation and steatosis. Histological analysis further indicated that SIM reduced lipid deposition in adipocytes and hepatocytes in comparison with that of the HFD group. The underlying mechanisms of SIM administration against HFD-induced hyperlipidemia were also studied by UPLC-Q-TOF/MS-based liver metabonomics coupled with pathway analysis. Metabolic pathway enrichment analysis of liver metabolites with significant difference in abundance indicated that fatty acids metabolism and amino acid metabolism were the main metabolic pathways altered by SIM administration. Meanwhile, operational taxonomic units (OTUs) analysis revealed that oral administration of SIM altered the composition of gut microbiota, including Ruminococcaceae (OTU960) and Lactobacillus (OTU152), and so on. Furthermore, SIM treatment also regulated the mRNA levels of the genes involved in lipid and cholesterol metabolism. Immunohistochemistry (IHC) analysis of the liver-related proteins (CD36, CYP7A1 and SREBP-1C) showed that oral administration of SIM could regulate the levels of the protein expression related to hepatic lipid metabolism.

Compound CYD19 has been identified as a CBP/p300-Snail interaction inhibitor that exhibits potent anticancer efficacy., The zinc finger transcription factor Snail is aberrantly activated in many human cancers and associated with poor prognosis. Therefore, targeting Snail is expected to exert therapeutic benefit in patients with cancer. However, Snail has traditionally been considered “undruggable,” and no effective pharmacological inhibitors have been identified. Here, we found a small-molecule compound CYD19 that forms a high-affinity interaction with the evolutionarily conserved arginine-174 pocket of Snail protein. In aggressive cancer cells, CYD19 binds to Snail and thus disrupts Snail’s interaction with CREB-binding protein (CBP)/p300, which consequently impairs CBP/p300-mediated Snail acetylation and then promotes its degradation through the ubiquitin-proteasome pathway. Moreover, CYD19 restores Snail-dependent repression of wild-type p53, thus reducing tumor growth and survival in vitro and in vivo. In addition, CYD19 reverses Snail-mediated epithelial-mesenchymal transition (EMT) and impairs EMT-associated tumor invasion and metastasis. Our findings demonstrate that pharmacologically targeting Snail by CYD19 may exert potent therapeutic effects in patients with cancer.

The Hippo-YAP signaling pathway plays an essential role in epithelial cells during intestinal regeneration and tumorigenesis. However, the molecular mechanism linking stromal signals to YAP-mediated intestinal regeneration and tumorigenesis is poorly defined. Here, we report a stroma-epithelium ISLR-YAP signaling axis essential for stromal cells to modulate epithelial cell growth during intestinal regeneration and tumorigenesis. Specifically, upon inflammation and in cancer, an oncogenic transcription factor ETS1 in stromal cells induces expression of a secreted protein ISLR that can inhibit Hippo signaling and activate YAP in epithelial cells. Deletion of Islr in stromal cells in mice markedly impaired intestinal regeneration and suppressed tumorigenesis in the colon. Moreover, the expression of stromal cell-specific ISLR and ETS1 significantly increased in inflamed mucosa of human IBD patients and in human colorectal adenocarcinoma, accounting for the epithelial YAP hyperactivation. Collectively, our findings provide new insights into the signaling crosstalk between stroma and epithelium during tissue regeneration and tumorigenesis.

Lactobacillus paracasei FZU103, a probiotic previously isolated from the traditional brewing process of Hongqu rice wine, may have the beneficial effect of improving the disorder of lipid metabolism. This study aimed to determine the beneficial effects of L. paracasei FZU103 on improving hepatic lipid accumulation associated with hyperlipidemia. Results indicated that L. paracasei FZU103 intervention significantly inhibited the abnormal growth of body weight and epididymal white adipose tissue (eWAT), prevented the hypertrophy of epididymal adipocytes, ameliorated the biochemical parameters of serum and liver related to lipid metabolism in HFD-fed mice. Histological analysis also showed that the excessive accumulation oflipid dropletsin the livers induced by HFD-feeding was greatly alleviated by L. paracasei FZU103 intervention. In addition, L. paracasei FZU103 also promoted the excretion of bile acids (BAs) through feces. Metagenomic analysis revealed that oral supplementation with L. paracasei FZU103 significantly increased the relative abundance of Ruminococcus, Alistipes, Pseudoflavonifractor and Helicobacter, but decreased the levels of Blautia, Staphylococcos and Tannerella in HFD-fed mice. The relationships between lipid metabolic parameters and intestinal microbial phylotypes were also revealed by correlation heatmap and network. Furthermore, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS)-based liver metabolomics demonstrated that L. paracasei FZU103 had a significant regulatory effect on the metabolic pathways of glycerophospholipid metabolism, fatty acid degradation, fatty acid elongation, unsaturated fatty acids biosynthesis, riboflavin metabolism, glycerolipid metabolism, primary bile acid biosynthesis, arachidonic acid metabolism, etc. Additionally, L. paracasei FZU103 intervention regulated expression of hepatic genes involved in lipid metabolism and bile acid homeostasis, and promoted fecal excretion of intestinal BAs. These findings present new evidence supporting that L. paracasei FZU103 has the potential to improve lipid metabolism, and could be used as a potential functional food for the prevention of hyperlipidemia.

This study aimed to investigate the effects of 55% ethanol extracts from G. lucidum (GL55) on high-fat diet (HFD) induced hyperlipidaemia and gut microbiota modulation in rats. UPLC-QTOF MS-MS indicated that GL55 was enriched with triterpenoids. GL55 markedly alleviated the dyslipidemia in HFD fed rats through decreasing the levels of serum total triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), alanine transaminase (ALT), free fatty acids (FFA), fasting blood-glucose (FBG) and inhibiting hepatic steatosis. Furthermore, high throughput sequencing analysis revealed that GL55 supplementation altered the composition of gut microbiota, in particular increased the relative abundance of functionally relevant enterotypes, such as Alloprevotella, Ruminococcus and Prevotella, but decreased the proportion of Turicibacter and Clostridium XVIII compared with the HFD group. The Spearman's correlation analysis revealed that Alloprevotella, Ruminococcus and Prevotella were negatively correlated with serum lipid profiles. Meanwhile, GL55 treatment regulated mRNA and protein expressions levels involved in lipid and glucose metabolism.

Hong Qu, which mainly contains Monascus sp. and other microorganisms, as well as numerous microbial metabolites, is used as the fermentation starter of Hong Qu glutinous rice wine, a traditional alcoholic beverage. Two widely-used types of Hong Qu, namely Wuyi Qu (WYQ) and Gutian Qu (GTQ), were thoroughly compared for their fermentation properties, volatile profiles, and microbiota structures in this study. Significantly higher color value, glucoamylase and α-amylase activities were discovered in WYQ. And substantial variation in volatile components and microbial communities were also observed between them. It was identified that bacterial genus Burkholderia dominated GTQ (71.62%) and Bacillus dominated WYQ (44.73%), while Monascus purpureus was the most abundant fungal species in both types of starters (76.99%). In addition, 213 bacterial genera and 150 fungal species with low-abundance were also detected. Since the Linear Discriminant Analysis Effect Size algorithm, 14 genus-level bacterial taxa and 10 species-level fungal taxa could be utilized to distinguish these two types of starters. Moreover, the potential correlation of the volatile components and microbiota within WYQ and GTQ were further analyzed, by utilizing Partial Least Squares Discriminant Analysis. Ultimately, this study provides detailed insight into the volatile profiles and microbial communities presented in Hong Qu.

Summary Androgenetic haploid embryonic stem cells (AG-haESCs) hold great promise for exploring gene functions and generating gene-edited semi-cloned (SC) mice. However, the high incidence of self-diploidization and low efficiency of SC mouse production are major obstacles preventing widespread use of these cells. Moreover, although SC mice generation could be greatly improved by knocking out the differentially methylated regions of two imprinted genes, 50% of the SC mice did not survive into adulthood. Here, we found that the genome-wide DNA methylation level in AG-haESCs is extremely low. Subsequently, downregulation of both de novo methyltransferase Dnmt3b and other methylation-related genes was determined to be responsible for DNA hypomethylation. We further demonstrated that ectopic expression of Dnmt3b in AG-haESCs could effectively improve DNA methylation level, and the high incidence of self-diploidization could be markedly rescued. More importantly, the developmental potential of SC embryos was improved, and most SC mice could survive into adulthood., Graphical Abstract, Highlights • Dnmt3b restores methylation of hypomethylated AG-haESCs • Dnmt3b suppresses self-diploidization of AG-haESCs • Dnmt3b promotes developmental potential of SC embryos • Dnmt3b improves SC mice survival to adulthood, Ectopic expression of Dnmt3b could rescue DNA methylation level in repetitive sequences of hypomethylated AG-haESCs, suppress high incidence of self-diploidization, and promote developmental potential of SC embryos, and most SC mice could survive into adulthood.

This study aimed to investigate the effects of 95% ethanol extract of G. frondosa (GF95) on lipid metabolism and gut microbiota composition in high-fat diet (HFD) fed rats. UPLC/Q-TOF MS indicated that GF95 was enriched with flavones, fatty acids and so on. Meanwhile, we found that body weight, serum lipid or liver index (total cholesterol, triglyceride, and low density lipoprotein cholesterol) levels were significantly decreased in GF95-treated HFD-fed rats. Furthermore, GF95 treatment regulated mRNA expression levels involved in lipid metabolism. GF95 consumption significantly enhanced the excretion of bile acids in the cecum. Besides, in this study, a higher abundance of Butyricimonas genus was revealed in the GF95 group, which is highly related to the highest production of short-chain fatty acids in the caecum contents among the experimental groups. Interestingly, results from network analysis showed that Butyricimonas were negatively correlated with serum and liver lipid profiles.

The contamination of peanuts with Aspergillus flavus and subsequent aflatoxins is considered to be one of the most serious safety problems in the world. Water activity (aw) and temperature are limiting factors for fungal growth and aflatoxins production during storage. To optimize the practical storage parameter, the effect of aw (0.85–0.99) and temperature (15–42 °C) on fungal growth, aflatoxin production and the expression of aflatoxin biosynthetic and regulatory genes in shelled peanuts was investigated. A. flavus grew at a lower rate when temperature ≤20 °C or aw ≤ 0.85. For the growth of A. flavus in shelled peanuts, the optimum conditions were aw was 0.98, and the optimum temperature was 37 °C. The maximum amount of AFB1 in peanuts was obtained at 28 °C and aw 0.96. Real-time analysis showed that 16 of 25 genes had highest expression levels at 28 °C under aw 0.92, while 9 genes had highest expression levels at 37 °C under aw 0.92. Compared with 37 °C, all aflatoxin biosynthetic pathway genes were down-regulated at 42 °C. All the pathway genes and laeA were up-expressed at aw of 0.96 under 28 °C, compared to aw 0.99. Furthermore, there was a good positive correlation between the ratio of aflS/aflR and AFB1 production. The expression of laeA was also positively correlated with AFB1 production while the expression of brlA was correlated with the A. flavus growth. The results of this study suggest that AFB1 production in peanut kernels can occur over a wider range of aw × temperatures levels compared to formula media and peanut media. Previous studies have showed that AFB1 could not be produced on formula media at 37 °C without the expression of most aflatoxin structural genes. But, in the un-autoclaved shelled peanuts, high concentration of AFB1 was produced at 37 °C with up-regulation of some aflatoxin biosynthetic genes. From a food safety point of view, the results can be used to optimize certain food technological processes and develop prevention strategies to control such carcinogenic natural metabolites in grains (such as peanuts, maize and rice) and derived products.

Due to the hindrance of flocculated polymers and bacterial cell wall, the production of Bacillus subtilis using monosodium glutamate byproduct (MSGB) was low. With the assistance of scanning electron microscope images, effects of alkali, lysozyme, papain, ultrasound, and their combinations on MSGB were evaluated using the results of soluble protein, carbohydrate, monosaccharides and peptidoglycans. Alkali could dissolve flocculated polymers increasing 21% soluble MSGB, and thus enhanced the subsequent treatments (ultrasound, lysozyme, or papain) to increase 14–17% soluble MSGB. As ultrasound mainly released intercellular components (mannose, and glucose) while lysozyme or papain mainly released cell wall components (peptidoglycans), the combination of alkali, ultrasound, and enzymes led to a highest soluble MSGB (78%), yielding a maximal B. subtilis production of 6.6 × 109 colony-forming units mL−1. This yield was about 33 times that of using untreated MSGB, and the key to improve B. subtilis production was the release of carbohydrate.

MicroRNA-mediated post-transcriptional regulation plays key roles in stem cell self-renewal and tumorigenesis. However, the in vivo functions of specific microRNAs in controlling mammary stem cell (MaSC) activity and breast cancer formation remain poorly understood. Here we show that miR-31 is highly expressed in MaSC-enriched mammary basal cell population and in mammary tumors, and is regulated by NF-κB signaling. We demonstrate that miR-31 promotes mammary epithelial proliferation and MaSC expansion at the expense of differentiation in vivo. Loss of miR-31 compromises mammary tumor growth, reduces the number of cancer stem cells, as well as decreases tumor-initiating ability and metastasis to the lung, supporting its pro-oncogenic function. MiR-31 modulates multiple signaling pathways, including Prlr/Stat5, TGFβ and Wnt/β-catenin. Particularly, it activates Wnt/β-catenin signaling by directly targeting Wnt antagonists, including Dkk1. Importantly, Dkk1 overexpression partially rescues miR31-induced mammary defects. Together, these findings identify miR-31 as the key regulator of MaSC activity and breast tumorigenesis.

Teramethylpyrazine (TMP) is a pyrazine alkaloid with a variety of pharmacological effects and widely used in clinical practice. In this study, Monascus strain M-3 with high TMP and low citrinin productions was screened from red yeast rice collected from different areas of Fujian province in China. Monascus strain M-3 was further identified through molecular biological methods based on the sequencing of different gene regions. Results showed that primer set of β-tubulin-F/β-tubulin-R targeting the β-tubulin gene was more suitable for the identification of Monascus strain M-3. Based on comparative analysis of the results obtained by different primer sets, Monacus strain M-3 was finally identified as Monascus purpureus. Then, the composition of culture medium and liquid state fermentation conditions of Monacus strain M-3 were optimized by single-factor experiment and central composite design of response surface methodology, with the purpose of optimizing the TMP productions. The optimum composition of culture medium and liquid state fermentation conditions are as follows: starch 5.1%, peptone 3.51%, Mg2SO4 0.1%, NaNO3 0.2%, KH2PO4 0.25%, fermentation temperature of 28.23 °C, fermentation time of 9 d, shaker speed of 180 r/min, inoculum size of 7.21% and loading volume of 100 mL/250 mL triangular flask. Under the optimum composition of culture medium and liquid state fermentation conditions, the TMP production of Monacus strain M-3 can reach 13.49 µg/mL. This study is expected to provide a new approach of TMP production through biosynthesis by Monascus strain, and promote the further development of functional foods or medicines from Monascus fermentation products.

High-fat diet (HFD) and sucrose intake can lead to hyperlipidemia, hypercholesterolemia, and nonalcoholic fatty liver disease (NAFLD) as well as disturbed gastrointestinal microbiota and dysfunctional intestinal barrier. In the present study, we showed that Ganoderma lucidum polysaccharide and chitosan (PC) significantly mitigated the hyperlipidemia in HFD-fed hamsters via lowering the contents of serum total triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and aspartate aminotransferase (AST). Furthermore, PC changed the composition of gastrointestinal microbiota and elevated the relative abundances of beneficial bacteria, such as Prevotella, Oscillibacter, and SCFA-producers. Interestingly, we also found that the abundances of Prevotella, Alloprevotella, Bifidobacterium, and Alistipes were negatively associated with serum lipid profiles. Collectively, the above-mentioned findings indicated that PC could improve lipid metabolic disorders, at least in part, by modulating gastrointestinal microbiota, suggesting that PC could be used as a potential lipid-lowering ingredient in functional foods. PRACTICAL APPLICATIONS: PC could ameliorate lipid metabolism disorder, at least in part, by regulating specific gut microbiota, suggesting its potential as a novel lipid-lowering ingredient in functional foods. We believed that our findings could be of interest to the readers because they help others further understand the gut microbiota alterations that occurred after PC supplementation in the context of metabolic syndrome (MetS).

This study aimed to investigate the hypoglycemic and hypolipidemic activities of polysaccharides from Grifola frondosa (GFP) in diabetic mice induced by high-fat diet (HFD) and streptozotocin (STZ). Results showed that oral administration of GFP markedly reduced the serum levels of fasting blood glucose (FBG), oral glucose tolerance (OGT), cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C), and significantly decreased the hepatic levels of TC, TG and free fatty acids (FFA). Meanwhile, high-dose of GFP supplementation (900 mg/kg day) also showed powerful effects on moderating the composition of intestinal microflora in diabetic mice, especially altering the functionally relevant intestinal microbial phylotypes. Spearman’s correlation network analysis revealed that key microbial phylotypes responding to GFP intervention were strongly correlated with the glucose and lipid metabolic disorders associated parameters. Moreover, GFP treatment regulated mRNA expression levels of the genes responsible for hepatic glucose and lipid metabolism. It is noteworthy that GFP treatment markedly increased mRNA expression of cholesterol 7α-hydroxylase (CYP7A1) and bile salt export pump (BSEP), suggesting an enhancement of bile acids (BAs) synthesis and excretion in liver. These findings demonstrated that GFP could prevent hyperglycemia and hyperlipidemia in diabetic mice by altering gut microbiota and regulating hepatic glycolipid metabolism related genes, and therefore could be used as potential functional food ingredients for the prevention or treatment of hyperglycemia and hyperlipidemia.

This study aimed to compare the hypolipidaemic activities of different Monascus pigments (yellow, red and orange pigments) and elucidate their possible regulatory mechanisms on lipid and cholesterol metabolism in rats fed on a high-fat diet (HFD). Results showed that oral administrations of Monascus yellow, red and orange pigments can markedly alleviate the disturbance of lipid metabolism through ameliorating the serum lipid levels and suppressing hepatic lipid accumulation and steatosis. Meanwhile, the excretion of fecal cholesterol, triacylglycerols and bile acids was also promoted by the oral administrations of different Monascus pigments (MPs). Furthermore, Monascus pigment (MP) supplementation produced significant structural changes in the intestinal microbiota of HFD-fed rats, and modulated the relative abundance of functionally related microbial phylotypes compared with the HFD group in particular. Key phylotypes in response to the HFD and Monascus pigment (MP) intervention were found to strongly correlate with the lipid metabolism disorder associated parameters using Spearman's correlation coefficient. Some beneficial gut microbiota (such as Oscillibacter sp., Ruminococcus albus, Clostridium sp., etc.) were found to be negatively correlated with the serum and hepatic lipid indicator. Moreover, Monascus pigment (MP) treatments regulated the mRNA expression levels of the genes responsible for lipid and cholesterol metabolism. In general, different Monascus pigments (MPs) regulate the homeostasis of lipid and cholesterol metabolism through different regulatory pathways. These findings illustrated that not only Monascus yellow pigments, but also Monascus red and orange pigments have the potential to ameliorate lipid metabolic disorders, and therefore could be used as potential functional food ingredients for the prevention or treatment of hyperlipidemia and gut microbiota dysbiosis.

Xuecai is a Chinese-fermented vegetable product that has attracted much attention because of its unique umami taste. However, investigations of the bacterial communities, umami components and chemical characteristics in xuecai and the brine have not yet been conducted. In this study, the bacterial communities and their correlations with the umami components and the chemical characteristics are investigated. The results showed that significantly higher contents of total protein, total acid, amino acid nitrogen, total sugar, titratable acidity, salt, amino acids and organic acids, as well as significantly lower nitrite concentrations were observed in the brine samples. The supervised pattern recognition method revealed that the xuecai and brine could be distinguished by these indicators. In addition, the xuecai and brine were both dominated by Lactobacillus, Halanaerobium and Halomonas. A correlation analysis showed that halophilic bacteria had a positive effect on the umami components and chemical characteristics of the xuecai and brine. However, lactic acid bacteria both showed a negative correlation with amino acids and most organic acids. Membrane transport, carbohydrate metabolism and amino acid metabolism were the most abundant pathways. These results enhance our understanding of the bacterial communities, as well as the umami components and chemical characteristics in Zhejiang xuecai and brine.

The main goal of this work was to investigate the bacterial flora present in the traditional fermentation starters using matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) mass spectrometry fingerprinting combined with 16S rRNA gene sequencing and species-specific PCRs. A total of 1314 bacterial strains were obtained from ten kinds of representative wine fermentation starters (5 Hong Qu and 5 Yao Qu) and classified by MALDI-TOF mass spectrometry. Sixteen different bacterial species belonging to six different genera (Bacillus, Staphylococcus, Leuconostoc, Pediococcus, Lactobacillus and Lactococcus) were identified with high accuracy by comparing them with generated spectra of MALDI Biotyper database. Comparison of 16S rRNA gene phylogeny and MALDI-TOF mass spectrometry analysis showed that MALDI-TOF mass spectrometry fingerprinting was a good complementary approach to 16S rRNA sequencing and even a more powerful tool in the accurate differentiation and classification of Bacillus species, especially for differentiating between Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus methylotrophicus. MALDI-TOF mass spectrometry was also found to provide valuable information at species levels for the genus of Lactobacillus. The identification revealed that most strains isolated from traditional fermentation starters were B. amyloliquefaciens (31.81%), followed by Bacillus megaterium (16.89%), Lc. lactis subsp. lactis (8.60%) and B. subtilis (8.14%). In the culture-dependent quantitative analysis, 69.01% of the detected isolates were Bacillus species, and 30.44% of the detected isolates belonged to the class of lactic acid bacteria (LAB), including Leu. mesenteroides, Pediococcus pentosaceus, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus brevis, Pediococcus acidilactici, Lactobacillus coryniformis and Lc. lactis subsp. lactis. This study provides detailed evaluation of bacterial communities in the traditional fermentation starters for Hong Qu glutinous rice wine using a fast and cost-effective technique—MALDI-TOF mass spectrometry fingerprinting. Information provided by this study is fundamental and useful to develop effective strategies for the selection of beneficial bacterial strains and improve of quality of Hong Qu glutinous rice wine.

The main disadvantage of DNA-based quantification methods is their inability to distinguish between viable and non-viable cells, as dead cells may also retain a significant amount of DNA. To overcome this limitation, propidium monoazide (PMA) treatment was applied to effectively prevent amplification of DNA from dead cells. The objectives of this study were to develop and evaluate an accurate detection method that combines PMA staining with real-time qPCR to quantify viable dominant microorganisms during the traditional brewing of Hong Qu glutinous rice wine. Firstly, three methodological factors (concentration of PMA, incubation time and photoactivation time) were optimized for representative strains of different microbial species. For the tested microbial strains ( Monascus purpureus , Saccharomyces cerevisiae and Lactobacillus plantarum ), a treatment of 25 μM PMA for 20 min incubation followed by 10 min photoactivation was considered optimal to achieve a compromise between minimal impact on intact cells and maximal signal reduction for the compromised cells. Compared with qPCR without PMA treatment, results of PMA-qPCR showed that the optimized PMA pretreatment can effectively exclude the dead microbial cells without any negative effects on the intact cells. The concentrations of viable cells calculated by PMA-qPCR were comparable (the same order of magnitude was obtained) with those obtained by culture-based method. Finally, the developed PMA-qPCR methods were successfully applied to monitor the total bacteria, LAB, fungi, yeasts, M. purpureus , S. cerevisiae and L. plantarum during the traditional brewing of Hong Qu glutinous rice wine. Both of the total bacterial and fungal populations increased significantly at the early brewing stage, but they decreased at the later brewing stage. PMA-qPCR results revealed that the predominant microorganisms varied in different brewing phases. L. plantarum and S. cerevisiae grew significantly and became the predominant bacterial and fungal species at the final stage of traditional brewing process, respectively. This study standardized the PMA-qPCR parameters for dominant microorganisms associated with the traditional brewing of Hong Qu glutinous rice wine, which would show great value in scientifically understanding of the brewing mechanism and provide useful information for the selection of beneficial microbial strains to improve wine quality.

In this study, reverse transcription quantitative real-time PCR (RT-qPCR) assays using species-specific primers were developed for the quantification of the three major fungal species (Saccharomycopsis fibuligera, Rhizopus oryzae, and Monascus purpureus) during the traditional brewing of Hong Qu glutinous rice wine. Species-specific primers (Sfi-F/Sfi-R, Ro-F/Ro-R, and Mp-F/Mp-R) targeting the ITS-5.8S rRNA gene and the beta-tubulin gene sequences were designed and their specificity was evaluated by RT-qPCR with the total RNA from fungal species closely related to the traditional brewing of Hong Qu glutinous rice wine. Results of RT-qPCR using species-specific primers showed 100 % inclusivity (positive signal in the presence of target complementary DNA (cDNA)) and 100 % exclusivity (negative signals in the presence of non-target template). The specificity of each primer set was also tested when the target cDNA was mixed with non-target cDNA, and results showed that there was no significant difference (P > 0.05) in threshold cycle (Ct) values obtained from pure target cDNA and target cDNA mixed with non-target cDNA. The detection limits of the established qPCR assays were determined to be 101 gene copies/μL for S. fibuligera and 102 gene copies/μL for R. oryzae and M. purpureus, respectively. Good correlations were obtained for the tested species between 102 and 108 gene copies/μL by qPCR analysis. Finally, the established RT-qPCR assays were applied to quantify viable S. fibuligera, R. oryzae, and M. purpureus during the traditional brewing of Hong Qu glutinous rice wine. This study standardized the RT-qPCR assays for dominant fungi associated with the traditional brewing of Hong Qu glutinous rice wine, and it also proved the usefulness of RT-qPCR assays for the quantification of live cells in multi-species samples. Results would show great value in scientifically understanding of the brewing mechanism of Hong Qu glutinous rice wine.

The aim of this study was to investigate the bacterial and fungal community structure and dynamics during the traditional brewing of Fuzhou Hong Qu glutinous rice wine through culture-dependent and culture-independent methods. The culture-independent DGGE profiles revealed the presence of eleven bacterial species (Janthinobacterium lividum, Lactobacillus plantarum, Weissella soli, Leuconostoc mesenteroides, Lactobacillus brevis, Pediococcus pentosaceus, Lactococcus lactis subsp. lactis, Pediococcus acidilactici, Bacillus subtilis, Bacillus aryabhattai and Bacillus megaterium) and seven fungal species (Saccharomycopsis fibuligera, Pichia guilliermondii, Saccharomyces cerevisiae, Wickerhamomyces anomalus, Rhizopus oryzae, Candida glabrata and Monascus purpureus) during the traditional brewing process. Furthermore, the microbial compositions of fermented samples were investigated using culture-dependent approach (conventional plate count technique combined with molecular identification). Results showed that the relative proportions of some fungal species (C. glabrata, W. anomalus, P. guilliermondii and S. fibuligera) and bacterial species (Lc. lactis subsp. lactis, Staphylococcus pasteuri, Bacillus sp., P. pentosaceus) detected at the early brewing stage decreased as the fermentation progressed, while S. cerevisiae, L. brevis, L. plantarum and Lactobacillus paracasei became the predominant species during the late brewing period. Some differences were also found between the profiles obtained by PCR–DGGE and culture-dependent technique. Comparative analysis revealed that some species (J. lividum, W. soli, B. aryabhattai and P. guilliermondii) could only be detected using PCR–DGGE technique. Conversely, some species (L. paracasei, S. pasteuri, Bacillus methylotrophicus, Mucor indicus and Aspergillus flavus) could only be detected by culture-dependent method. Therefore, the combination of culture-dependent and -independent approaches is quite necessary because they provided complementary information about the composition of the microbial community during the traditional brewing of Fuzhou Hong Qu glutinous rice wine. This study provides the first detailed evaluation of fungal community and dynamics in Fuzhou Hong Qu glutinous rice wine traditional brewing process using culture-dependent and -independent methods, which would facilitate scientific understanding of the traditional brewing mechanism and be useful for the selection of the suitable and beneficial strains to improve the wine quality.

Monascus-fermented rice has been used to treat digestive disorder and promote blood circulation in China and other Asian countries for centuries. However, the effects and mechanisms of Monascus purpureus-fermented common buckwheat (HQ) on non-alcoholic fatty liver disease (NAFLD) and dyslipidemia are unclear. Here, oral supplementation of HQ significantly inhibited the abnormal growth of body weight and epididymal white adipose tissue (eWAT), prevented the hypertrophy of epididymal adipocytes, ameliorated some biochemical parameters of serum and liver related to lipid metabolism in mice fed a high-fat and high-cholesterol diet (HFD). Histological analysis also showed that the excessive accumulation of lipid droplets in the livers induced by HFD-feeding was greatly alleviated by HQ supplementation. Metagenomic analysis revealed that HQ supplementation made significant structural changes in the intestinal microflora of mice fed with HFD. The Spearman's correlation analysis revealed that physiological index, serum and liver lipid profiles were positively correlated with Bacteroidales S24-7, Streptococcus, Allobaculum, and Clostridiales XIII, but negatively associated with Lactobacillus, Ruminococcaceae_NK4A214 group, Ruminiclostridium, and Alistipes. UPLC-QTOF/MS-based liver metabolomics demonstrated that HQ intervention had significant regulatory effects on the metabolic pathways of primary bile acid biosynthesis, pyrimidine metabolism, ether lipid metabolism, glutathione metabolism, glycine, serine and threonine metabolism, and amino sugar and nucleotide sugar metabolism, etc. Additionally, HQ intervention regulated the mRNA levels of hepatic genes involved in hepatic lipid metabolism and bile acid homeostasis. Collectively, these findings present new evidence supporting that HQ has the potential to ameliorate dyslipidemia and NAFLD via modulating the intestinal microbial populations and hepatic metabolite profile in hyperlipidemic mice induced by HFD.

Monascus vinegar is one of the most famous and popular Chinese vinegars. The present study identified 60 volatile compounds, 23 amino acids, and seven organic acids during the traditional brewing of Monascus vinegar. Acetic acid, alanine, alcohols, esters, lactic acid, and valine were the predominant metabolic compounds found during the fermentation process. Komagataeibacter medellinensis, Lactobacillus acetotolerans, Saccharomycopsis fibuligera, Sterigmatomyces halophilus, and Yarrowia lipolytica were the dominant microorganisms during the traditional brewing of Monascus vinegar. Furthermore, based on Spearman's correlation analysis, K. medellinensis showed a positive correlation with acetic acid, acetoin, benzaldehyde, phenethyl acetate, 4-ethylphenol, proline, threonine, and isoleucine. Saccharomyces cerevisiae was positively associated with the production of acetoin, benzeneacetaldehyde, 2,3,5-trimethylpyrazine, proline, threonine, and isoleucine. Bacillus velezensis and Yarrowia lipolytica were positively correlated with esters and alcohols, implying that these microorganisms might make a significant contribution to the flavor of vinegar. These findings demonstrated that some microorganisms may play important roles in improving the aromatic quality of Monascus vinegar.

Accumulating evidence indicates that the zinc-finger transcription factor ZEB1 is predominantly expressed in the stroma of several tumours. However, the role of stromal ZEB1 in tumour progression remains unexplored. In this study, while interrogating human databases, we uncover a remarkable decrease in relapse-free survival of breast cancer patients expressing high ZEB1 levels in the stroma. Using a mouse model of breast cancer, we show that ZEB1 inactivation in stromal fibroblasts suppresses tumour initiation, progression and metastasis. We associate this with reduced extracellular matrix remodeling, immune cell infiltration and decreased angiogenesis. ZEB1 deletion in stromal fibroblasts increases acetylation, expression and recruitment of p53 to FGF2/7, VEGF and IL6 promoters, thereby reducing their production and secretion into the surrounding stroma. Importantly, p53 ablation in ZEB1 stroma-deleted mammary tumours sufficiently recovers the impaired cancer growth and progression. Our findings identify the ZEB1/p53 axis as a stroma-specific signaling pathway that promotes mammary epithelial tumours., In epithelial cells Zeb1 is involved in the epithelial to mesenchymal transition. In this study, the authors show in a mouse model of breast cancer, that Zeb1 expression in stromal cells is required for tumour formation and metastasis.

High mobility group box-1 (HMGB1) could function as an early trigger for pro-inflammatory activation after spinal cord injury (SCI). Spinal cord edema contributes to inflammatory response mechanisms and a poor clinical prognosis after SCI, for which efficient therapies targeting the specific molecules involved remain limited. This study was designed to evaluate the roles of HMGB1 on the regulation of early spinal cord edema, astrocyte activation, and aquaporin-4 (AQP4) expression in a rat SCI model. Adult female Sprague-Dawley rats underwent laminectomy at T10, and the SCI model was induced by a heavy falling object. After SCI, rats received ethyl pyruvate (EP) or glycyrrhizin (GL) via an intraperitoneal injection to inhibit HMGB1. The effects of HMGB1 inhibition on the early spinal cord edema, astrocyte activation (glial fibrillary acidic protein [GFAP] expression), and AQP4 expression after SCI (12 h-3 days) were analyzed. The results showed that EP or GL effectively inhibited HMGB1 expression in the spinal cord and HMGB1 levels in the serum of SCI rats. HMGB1 inhibition improved motor function, reduced spinal cord water content, and attenuated spinal cord edema in SCI rats. HMGB1 inhibition decreased SCI-associated GFAP and AQP4 overexpression in the spinal cord. Further, HMGB1 inhibition also repressed the activation of the toll-like receptor 4/myeloid differentiation primary response gene 88/nuclear factor-kappa B signaling pathway. These results implicate that HMGB1 inhibition improved locomotor function and reduced early spinal cord edema, which was associated with a downregulation of astrocyte activation (GFAP expression) and AQP4 expression in SCI rats.

Aflatoxin B1 (AFB1), which is mainly produced by Aspergillus flavus and Aspergillus parasiticus, is the most toxic and hepatocarcinogenic polyketide known. Chemical fungicides are currently utilized to reduce this fungal contaminant, but they are potentially harmful to human health and the environment. Therefore, natural anti-aflatoxigenic products are used as sustainable alternatives to control food and feed contamination. For example, eugenol, presents in many essential oils, has been identified as an aflatoxin inhibitor. However, its exact mechanism of inhibition is yet to be clarified. In this study, the anti-aflatoxigenic mechanism of eugenol in A. flavus was determined using a comparative transcriptomic approach. Twenty of twenty-nine genes in the aflatoxin biosynthetic pathway were down-regulated by eugenol. The most strongly down-regulated gene was aflMa, followed by aflI, aflJ, aflCa, aflH, aflNa, aflE, aflG, aflM, aflD, and aflP. However, the expression of the regulator gene aflR did not change significantly and the expression of aflS was slightly up-regulated. The down-regulation of the global regulator gene veA resulted in the up-regulation of srrA, and the down-regulation of ap-1 and mtfA. The early developmental regulator brlA was profoundly up-regulated in A. flavus after eugenol treatment. These results suggested a model in which eugenol improves fungal development by up-regulating the expression of brlA by the suppression of veA expression and inhibits aflatoxin production through the suppression of veA expression. Exposure to eugenol also caused dysregulated transcript levels of the G protein-coupled receptors (GPCRs) and oxylipins genes. A Gene Ontology analysis indicated that the genes that were highly responsive to eugenol were mainly enriched in RNA-binding functions, suggesting that post-transcriptional modification plays a pivotal role in aflatoxin biosynthesis. KEGG analysis showed that ribosome biogenesis was the most dysregulated pathway, suggesting that eugenol dysregulates ribosome biogenesis, which then interrupts the biosynthesis of Nor-1, Ver-1, and OmtA, and prevents aflatoxisomes performing their normal function in aflatoxin production. In conclusion, our results indicated that eugenol inhibited AFB1 production by modulating the expression of structural genes in aflatoxin pathway, fungal antioxidant status, post-transcriptional modifications and biosynthesis of backbone enzymes in A. flavus.

The objective of this study was to explore the core functional microbiota for the production of volatile flavour during the traditional brewing of Wuyi Hong Qu glutinous rice wine, one of the most typical representatives of rice wine in China. Microbiological analysis based on high-throughput sequencing (HTS) technology demonstrated that bacteria of Lactobacillus, Bacillus, Leuconostoc, Lactococcus, Raoultella, Staphylococcus, Pediococcus, and Weissella, and fungi of Saccharomyces, Saccharomycopsis, Rhizopus, Monascus, Pichia, Wickerhamomyces, Candida, and Aspergillus were the predominant genera during the traditional fermentation process. Principal component analysis (PCA) based on the relative abundance showed that both of bacterial and fungal communities varied significantly in different fermentation phases. Some predominant microbial species or genera (including bacteria of Bacillus spp., Staphylococcus spp., Weissella spp., and P. acidilactici, and fungi of M. purpureus, R. oryzae, R. arrhizus var. arrhizus, and A. niger) were detected at the initial brewing stage, and their populations decreased as the fermentation progressed, while those of Lactobacillus, Gluconacetobacter, Leuconostoc, Pichia, Wickerhamomyces, and Saccharomyces increased to become the predominant genera at the final stage. A total of 79 volatile compounds were identified in traditional fermentation starters and during the traditional brewing process, mainly including esters, alcohols, acids, aldehydes, ketones, and phenols. Heatmaps and PCA also revealed the significant variances in the composition of volatile compounds among different samples. Furthermore, the potential correlations between microbiota succession and volatile flavour dynamics were explored through bidirectional orthogonal partial least squares (O2PLS) based correlation analysis. Three bacterial genera, namely, Gluconacetobacter, Lactobacillus, Lactococcus, and three fungal genera of Pichia, Wickerhamomyces, and Saccharomyces, were determined as the core functional microbiota for production of main volatile compounds in Wuyi Hong Qu glutinous rice wine. To conclude, information provided by this study is valuable to the development of effective strategies for the selection of beneficial bacterial and fungal strains to improve the quality of Wuyi Hong Qu glutinous rice wine.

Regulation of gene expression by epigenetic modifications such as DNA methylation is crucial for developmental and disease processes, including cell differentiation and cancer development. Genes repressed by DNA methylation can be derepressed by various compounds that target DNA methyltransferases, histone deacetylases, and other regulatory factors. However, some additional, unknown mechanisms that promote DNA methylation-mediated gene silencing may exist. Chemical agents that can counteract the effects of epigenetic repression that is not regulated by DNA methyltransferases or histone deacetylases therefore may be of research interest. Here, we report the results of a high-throughput screen using a 308,251-member chemical library to identify potent small molecules that derepress an EGFP reporter gene silenced by DNA methylation. Seven hit compounds were identified that did not directly target bulk DNA methylation or histone acetylation. Analyzing the effect of these compounds on endogenous gene expression, we discovered that three of these compounds (compounds LX-3, LX-4, and LX-5) selectively activate the p38 mitogen-activated protein kinase (MAPK) pathway and derepress a subset of endogenous genes repressed by DNA methylation. Selective agonists of the p38 pathway have been lacking, and our study now provides critical compounds for studying this pathway and p38 MAPK-targeted genes repressed by DNA methylation.