The progression from metabolic dysfunction-associated steatotic liver disease (MASLD) to steatohepatitis (MASH) entails rapid, often irreversible hepatic injury, underscoring the urgent need for innovative therapeutic strategies. Here, we demonstrate that excessive dietary sugar intake, particularly fructose, exacerbates liver disease progression through microbiota-mediated amplification of endogenous acetaldehyde production. Analysis of over 210,000 participants from the UK Biobank revealed a dose-dependent correlation between sugar consumption and liver-related mortality, accompanied by a microbial shift favoring acetaldehyde/ethanol fermentation pathways in MASH patients. Mechanistically, gut-derived acetaldehyde activates hepatic stellate cells via upregulation of matrix metalloproteinase-7 (MMP7), driving fibrogenesis. To mitigate this, we engineered Ligilactobacillus salivarius HAM, a probiotic strain with enhanced acetaldehyde-degrading capacity, which effectively halted fibrosis progression in preclinical models of diet-induced liver disease. These findings highlight microbiota-targeted modulation of aldehyde metabolism as a promising therapeutic avenue to intercept the transition from MASLD to MASH.

IntroductionBile BA profiling is important for understanding the pathogenetic mechanisms underlying gallbladder and gastrointestinal diseases. Until now, bile BAs have been explored mainly in adult male mice. However, sex- and age-related variations in the bile BA composition of normal experimental mice remain poorly characterized.MethodsBAs in bile from healthy, age-and sex-matched mice (8-week-old and 60-week-old) were quantified using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometer (UPLC-TQ-MS). The bile BA profiles were comprehensively compared across different ages and sexes, along with comparisons within the same sex at different ages.ResultsTotal BAs (TBAs) in female mice were higher than those in male mice at both ages, although total BAs and BA profiles were similar between the two ages. High concentrations of BAs contributed significantly to the observed sex differences at both ages. Notably, both high- and low-concentration BAs were regulated with increasing age in male mice, while only low-concentration BAs were modulated with age in female mice. A remarkable sex difference in total cholesterol (TC) was also observed, along with a significant negative association between TC and BA profiles.ConclusionThe findings demonstrate that bile BA profiles differ markedly with both sex and physiological age in commonly used normal laboratory mice, suggesting the need to consider for physiological differences related to sex and age when selecting suitable animals for pharmaceutical research and for mechanistic studies.

Background: Porcine bile powder (PBP) is a traditional Chinese medicine that has been used for centuries in various therapeutic applications. However, PBP has not previously undergone comprehensive component analysis and not been evaluated for safety through standard in vivo toxicological studies.Methods: In our study, we characterized the component of PBP by liquid chromatography-mass spectrometry. The acute and subchronic oral toxicity, genotoxicity, and teratogenicity studies of PBP were designed and conducted in Kunming mice and Sprague-Dawley (SD) rats.Results: The chemical analysis of PBP showed that the main components of PBP were bile acids (BAs), especially glycochenodeoxycholic acid. There were no signs of toxicity observed in the acute oral test and the subchronic test. In the genotoxicity tests, no positive results were observed in the bacterial reverse mutation test. Additionally, in the mammalian micronucleus test and mouse spermatocyte chromosomal aberration test, no abnormal chromosomes were observed. In the teratogenicity test, no abnormal fetal development was observed.Conclusion: Our findings demonstrate that PBP, composed mainly of BAs, is non-toxic and safe based on the conditions tested in this study.

建立了 一种基于超高效液相色谱单四极杆质谱检测器(UPLC-QDA)定量血浆游离氨基酸谱的方法.血浆样本经甲醇去除蛋白后,所得到的上清液挥干后用6-氨基喹啉-N-羟基琥珀酰亚胺基氨基甲酸酯(AQC)进行衍生.反应液在Cortecs C18色谱柱上进行6 min梯度洗脱达到色谱分离,采用正离子模式下的选择离子扫描模式测定.结果表明,34种氨基酸的检出限与定量限分别在0.1～5.0 μmol/L和0.2～10.0 μmol/L之间,并具有较好的线性关系(R2＞0.991)和良好的准确度(80.4％～121.8％),较高的精密度(相对标准偏差＜15％)以及较好的稳定性,低、中、高3个浓度加标回收率在78.1％～127.6％之间,基质效应在-21.9％～29.7％之间.本方法简单快速,稳定可靠,并成功地应用于正常人和小鼠血浆样本中氨基酸谱的定量.

Aging is a complex physiological process associated with degenerative disorder of metabolism and immune function, which contributes to the occurrence of senile diseases. The gut microbiota affects systemic inflammation in aging processes probably through metabolism, but their relationship is still unclear. In this study, 16S-rRNA-sequencing technology, gas chromatography-time-of-flight mass spectrometry (GC-TOFMS)–based metabolic profiling, and immune factor analysis combined with advanced differential and association analysis were employed to investigate the correlation between the microbiome, metabolome, and immune factors in male Wistar rats across lifespan. Our findings showed significant changes in the ileum microbiome and serum metabolome compositions across aging process. A two-level strategy was applied to demonstrate that key metabolites associated with age such as 4-hydroxyproline, proline, and lysine were clustered together and positively correlated with beneficial microbes including Bifidobacterium , Lactobacillus , and Akkermansia . Function analysis explored association between serum metabolite class and specific gut bacteria’s metabolism pathways. Further correlation analysis on all the alteration patterns provided an interaction network of main immune factors such as IL-10, IgA, IgM, and IgG with key gut bacteria and serum metabolites. This study offers new insights into the relationship between immune factors, serum metabolome, and the gut microbiome.

Calorie restriction (CR) can improve the metabolic balance of adults and elevate the relative abundance of probiotic bacteria in the gut while promoting longevity. However, the interaction between remodeled intestinal flora and metabolic improvement, as well as the mechanism for probiotic bacterial increase, are still unclear. In this study, using a metabolomics platform, we demonstrate for the first time, that CR leads to increased levels of malate and its related metabolites in biological samples. Next, we investigated the effects of CR on the gut microbial genome and the expression of mRNA related to energy metabolism which revealed a partially elevated TCA cycle and a subsequently promoted glyoxylate cycle, from which large amounts of malate can be produced to further impact malate related pathways in the host liver. Through the identification of key “hungry” metabolites produced by the gut microbiota that function in the promotion of autophagy in the host, further insight has been gained about a functional metabolic network important for both host-microbial symbiosis and maintenance of host health.### Competing Interest StatementThe authors have declared no competing interest.

Pu-erh tea displays cholesterol-lowering properties, but the underlying mechanism has not been elucidated. Theabrownin is one of the most active and abundant pigments in Pu-erh tea. Here, we show that theabrownin alters the gut microbiota in mice and humans, predominantly suppressing microbes associated with bile-salt hydrolase (BSH) activity. Theabrownin increases the levels of ileal conjugated bile acids (BAs) which, in turn, inhibit the intestinal FXR-FGF15 signaling pathway, resulting in increased hepatic production and fecal excretion of BAs, reduced hepatic cholesterol, and decreased lipogenesis. The inhibition of intestinal FXR-FGF15 signaling is accompanied by increased gene expression of enzymes in the alternative BA synthetic pathway, production of hepatic chenodeoxycholic acid, activation of hepatic FXR, and hepatic lipolysis. Our results shed light into the mechanisms behind the cholesterol- and lipid-lowering effects of Pu-erh tea, and suggest that decreased intestinal BSH microbes and/or decreased FXR-FGF15 signaling may be potential anti-hypercholesterolemia and anti-hyperlipidemia therapies.

There is increased appreciation for the diverse roles of the microbiome-gut-brain axis on mammalian growth and health throughout the lifespan. Numerous studies have demonstrated that the gut microbiome and their metabolites are extensively involved in the communication between brain and gut. Association study of brain metabolome and gut microbiome is an active field offering large amounts of information on the interaction of microbiome, brain and gut but data size and complicated hierarchical relationships were found to be major obstacles to the formation of significant, reproducible conclusions. This study addressed a two-level strategy of brain metabolome and gut microbiome association analysis of male Wistar rats in the process of growth, employing several analytical platforms and various bioinformatics methods. Trajectory analysis showed that the age-related brain metabolome and gut microbiome had similarity in overall alteration patterns. Four high taxonomical level correlated pairs of "metabolite type-bacterial phylum", including "lipids-Spirochaetes", "free fatty acids (FFAs)-Firmicutes", "bile acids (BAs)-Firmicutes", and "Neurotransmitters-Bacteroidetes", were screened out based on unit- and multivariant correlation analysis and function analysis. Four groups of specific "metabolite-bacterium" association pairs from within the above high level key pairs were further identified. The key correlation pairs were validated by an independent animal study. This two-level strategy is effective in identifying principal correlations in big data sets obtained from the systematic multiomics study, furthering our understanding on the lifelong connection between brain and gut.

Ginseng, a widely used functional food and food additive, has been proven to have promotion effects of health on the body. However, whether the long-term intake of Ginseng is beneficial or has side effects on an organism is still unclear. In this study, untargeted GC-TOFMS metabolomic analysis of serum, cecum and ileum intestinal contents was conducted to understand the effect of the long-term intake of Ginseng extracts. 16S rRNA microbial sequencing technology was applied to investigate the effect of Ginseng extracts on the structure of gut microbiota. Cytokines in spleen were detected to determine the effect of Ginseng extracts on the immune system. Compared to control groups, the metabolites in serum, cecum and ileum, such as amino acids, amines and other metabolites related to carbohydrate metabolism, significantly varied between the C and GS groups. Ginseng extracts affected the structure of gut microbiota with a decreased abundance of TM7, while the abundance of Proteobacteria, Methylobacteriaceae, Parasutterella, Sutterella increased in the GS group. The increased abundance of Bifidobacterium and Lactobacillus demonstrated that Ginseng extracts contribute to probiotic amplification. Highly correlated with Bifidobacterium and Lactobacillus, interleukin 4 (IL4), IL10 and immunoglobulin A (IgA) levels were significantly elevated after the long-term intake of Ginseng extracts. These results indicated that the long-term administration of Ginseng extracts positively affected the host-gut metabolism, immune system, the anti-inflammation process and the gut intestinal microbiota structure.

Different components of Panax ginseng have different properties and medicinal effects. Metabonomics was a prospective approach to analyze the global response of endogenous metabolites to physiological and pathological processes. In this study, an untargeted metabonomics method using GC/TOFMS combined with multivariate statistical techniques was applied to compare entire metabolite differences and the antistress variations among four components of P. ginseng, namely, total ginsenosides (TG), panaxadiol (PD), panaxatriol (PT), and ginseng polysaccharide (PS), in Wistar rats. The results of metabolite analysis showed that numerous urine metabolites involving neurotransmitters, amino acids, organic acids, and gut microbiota metabolites were changed after administration of the four components of P. ginseng, with TG having the least impact on urinary metabolites. The urinary metabolite profiling of these rats exposed to acute combined stress (forced swimming and behavior restriction) demonstrated that the four ginseng components attenuated urine metabolite changes involving gut microbiota metabolites, tricarboxylic acid (TCA) cycle and energy metabolites, and organic acids to different degrees, with TG improving most of the metabolites altered by stress.

Food withdrawal as a health-enhancing measure has beneficial effects on aging, disease prevention, and treatment. However, the cellular and molecular mechanisms involving gut microbial changes and metabolic consequences resulting from food withdrawal have yet to be elucidated. In this study, we subjected lean and obese mice to a dietary intervention that consisted of a 4-d complete food withdrawal and an 8-d 50% food withdrawal, and we studied changes in cecal microbiome and host serum metabolome. The abundance of potentially pathogenic Proteobacteria was decreased and Akkermansia muciniphila was elevated by food withdrawal in mice fed a high-fat diet (HFD). Meanwhile, food withdrawal decreased the abundance of metabolites in branched chain amino acid, lipid, and free fatty acid metabolisms in host serum, more so in HFD mice than in normal mice. Microbial predicted function also showed that food withdrawal decreased the abundance of microbes associated with predicted diseases in the HFD group but not in the normal chow group. Correlation between the microbiome data and metabolomics data revealed a strong association between gut microbial and host metabolic changes in response to food withdrawal. In summary, our results showed that food withdrawal was safer and more metabolically beneficial to HFD-induced obese mice than to normal lean mice, and the beneficial effects were primarily derived from the changes in gut microbiota, which were closely associated with the host metabolome.-Zheng, X., Zhou, K., Zhang, Y., Han, X., Zhao, A., Liu, J., Qu, C., Ge, K., Huang, F., Hernandez, B., Yu, H., Panee, J., Chen, T., Jia, W., Jia, W. Food withdrawal alters the gut microbiota and metabolome in mice.

Hyocholic acid (HCA) and its derivatives are found in only trace amounts in human blood, but constitute approximately 76 % of the bile acid (BA) pool in the pig, a species known for its exceptional resistance to type 2 diabetes mellitus (T2DM). Here we show that HCA species play a crucial role in maintaining glucose homeostasis and preventing T2DM. We found that in two cohort studies (n=1,213), both obesity and diabetes were associated with lower serum concentrations of HCA species. Serum HCA levels in apparently healthy individuals (n=132) were found to be strong predictors for metabolic health 10 years later. Oral administration of HCA increased serum fasting GLP-1, to a greater extent than metformin, in healthy and diabetic mouse models. HCA upregulated GLP-1 secretion in intestinal enteroendocrine cells via simultaneously activating G-protein-coupled BA receptor, TGR5, and inhibiting farnesoid X receptor, a unique mechanism that is not found in other BA species.

The advent of mass spectrometry-based analytical technologies coupled with multivariate statistical methods offer tremendous new opportunities for understanding the pharmacokinetics (PKs) of multicomponent herbal medicines (HMs). We recently proposed a poly-PK strategy to characterize the concentration-time profile and the metabolic response profile of multicomponent HMs using an integrated phytochemical and metabolomics approach. Here, we provided the first example of the poly-PK strategy, in which we simultaneously characterized the PK as well as the metabolic response profiles of a Chinese HM, Huangqi decoction (HQD, consisting of Radix Astragali and Radix Glycyrrhizae), in healthy Chinese volunteers. Using the poly-PK approach, we identified 56 HQD-derived compounds and 292 biotransformed HQD metabolites in human plasma. Additionally, we acquired the concentration-time profiles of these plasma HQD metabolites and correlated them with a plasma metabolomics profile consisting of 166 human endogenous metabolites that were significantly altered in response to HQD intervention.

Objective:Clinical biological samples are valuable resources for basic and clinical studies.It is urgent to develop a data information management system.Methods:Based on various advanced data processing and database technologies,we designed and developed an integrated information management system for clinical resource database.Results:Multi-source data from diverse clinical departments could be extracted,screened,associated and integrated.Such integrated information management system could provide a variety of foreground application and background management functions.Conclusion:This system is stable and easy to operate and is able to connect seamlessly to the sample storage system,the frozen storage equipment management system and the automatic workstation for sample processing.It can play an important role in the sample database management,data sharing and scientific research work.It was utilized as one of the common tools for the metabolic disease biobank of Shanghai No.6 People's Hospital.

NOS1AP variant rs12742393 is a functional single nucleotide polymorphism (SNP) and has been reported to be associated with schizophrenia and type 2 diabetes (T2DM) susceptibility in different populations. However, the molecular mechanisms are not clear. The main focus of the present study was to identify metabolic differences among different genotypes of the variant and to identify potential physiological and pathological mechanisms for the diseases.

Emerging evidence points to a strong association between sex and gut microbiota, bile acids (BAs), and gastrointestinal cancers. Here, we investigated the mechanistic link between microbiota and hepatocellular carcinogenesis using a streptozotocin-high fat diet (STZ-HFD) induced nonalcoholic steatohepatitis-hepatocellular carcinoma (NASH-HCC) murine model and compared results for both sexes. STZ-HFD feeding induced a much higher incidence of HCC in male mice with substantially increased intrahepatic retention of hydrophobic BAs and decreased hepatic expression of tumor-suppressive microRNAs. Metagenomic analysis showed differences in gut microbiota involved in BA metabolism between normal male and female mice, and such differences were amplified when mice of both sexes were exposed to STZ-HFD. Treating STZ-HFD male mice with 2% cholestyramine led to significant improvement of hepatic BA retention, tumor-suppressive microRNA expressions, microbial gut communities, and prevention of HCC. Additionally the sex-dependent differences in BA profiles in the murine model can be correlated to the differential BA profiles between men and women during the development of HCC. These results uncover distinct male and female profiles for gut microbiota, BAs, and microRNAs that may contribute to sex-based disparity in liver carcinogenesis, and suggest new possibilities for preventing and controlling human obesity-related gastrointestinal cancers that often exhibit sex differences.

Increasing evidences support that metabolically healthy obese (MHO) is a transient state. However, little is known about the early markers associated with the development of metabolic abnormalities in MHO individuals. Serum free fatty acids (FFAs) profile is highlighted in its association with obesity-related insulin resistance, type 2 diabetes mellitus (T2DM) and cardiovascular diseases (CVD). To examine the association of endogenous fatty acid metabolism with future development of metabolic abnormalities in MHO individuals, we retrospectively analyzed 24 [product FFA]/[precursor FFA] ratios in fasting sera and clinical data from 481 individuals who participated in three independent studies, including 131 metabolic healthy subjects who completed the 10-year longitudinal Shanghai Diabetes Study (SHDS), 312 subjects cross-sectionally sampled from the Shanghai Obesity Study (SHOS) and 38 subjects who completed an 8-week very low carbohydrate diet (VLCD) intervention study. Results showed that higher baseline level of oleic acid/stearic acid (OA/SA) and lower levels of stearic acid/palmitic acid (SA/PA) and arachidonic acid/dihomo-γ-linolenic acid (AA/DGLA) ratios were associated with higher rate of MHO to MUO conversion in the longitudinal SHDS. Further, the finding was validated in the cross-sectional and interventional studies. This panel of FFA ratios could be used for identification and early intervention of at-risk obese individuals.

转化医学研究是医学研究的新热点,资源整合和多学科协同是转化医学的特色,其对资源管理方式和信息挖掘技术有很高的依赖性和特殊需求.旨在针对转化医学研究的特点和通用流程,基于网络和数据库技术,对多款自主研发的资源管理软件进行模块化和后台化改造,依托ASP和Oracle技术将所有模块进行关联和组合,为转化医学研究人员提供一站式服务,实现生物样本、临床信息、检测仪器、存储设备、实验耗材、检测数据、研究项目、标准品和研究结果等科研资源的集中化和网络化管理,提高转化医学科研资源管理的集成化、标准化和自动化程度,提高多学科协同工作的效率和资源利用率,为转化医学研究提供技术支撑.

Comprehensive and accurate characterization of brain metabolome is fundamental to brain science, but has been hindered by technical limitations. We profiled the brain metabolome in male Wistar rats at different ages (day 1 to week 111) using high-sensitivity and high-resolution mass spectrometry. Totally 380 metabolites were identified and 232 of them were quantitated. Compared with anatomical regions, age had a greater effect on variations in the brain metabolome. Lipids, fatty acids and amino acids accounted for the largest proportions of the brain metabolome and their concentrations varied across the lifespan. The levels of polyunsaturated fatty acids were higher in infancy (week 1 to week 3) compared with later ages and the ratio of omega-6 to omega-3 fatty acids increased in the aged brain (week 56 to week 111). Importantly, a panel of 20 bile acids were quantitatively measured, most of which have not previously been documented in the brain metabolome. This study extends the breadth of the mammalian brain metabolome as well as our knowledge of functional brain development, both of which are critically important to move the brain science forward.

Glucocorticoids are commonly used in anti-inflammatory and immunomodulatory therapies, but glucocorticoid withdrawal can result in life-threatening risk of adrenal insufficiency. Chinese patented pharmaceutical product Jinkui Shenqi pill (JKSQ) has potent efficacy on clinical adrenal insufficiency resulting from glucocorticoid withdrawal. However, the underlying molecular mechanism remains unclear. We used an animal model to study JKSQ-induced metabolic changes under adrenal insufficiency and healthy conditions. Sprague-Dawley rats were treated with hydrocortisone for 7 days with or without 15 days of JKSQ pretreatment. Sera were collected after 72 h hydrocortisone withdrawal and used for global and free fatty acids (FFAs)-targeted metabolomics analyses using gas chromatography/time-of-flight mass spectrometry and ultraperformance liquid chromatography/quadruple time-of-flight mass spectrometry. Rats without hydrocortisone treatment were used as controls. JKSQ pretreatment normalized the significant changes of 13 serum metabolites in hydrocortisone-withdrawal rats, involving carbohydrates, lipids, and amino acids. The most prominent effect of JKSQ was on the changes of FFAs and some [product FFA]/[precursor FFA] ratios, which represent estimated desaturase and elongase activities. The opposite metabolic responses of JKSQ in adrenal insufficiency rats and normal rats highlighted the "Bian Zheng Lun Zhi" (treatment based on ZHENG differentiation) guideline of TCM and suggested that altered fatty acid metabolism was associated with adrenal insufficiency after glucocorticoid withdrawal and the protective effects of JKSQ.