Microbial succession and its effect on key aroma components during light-aroma-type Xiaoqu Baijiu brewing process.

Light-aroma-type Baijiu is a Chinese distilled alcoholic beverage produced from fermented sorghum. Microbial composition and dynamics during Baijiu production have a great influence on the flavor and quality of Chinese Baijiu. However, the microbial changes that occur during brewing of Xiaoqu Baijiu are poorly understood. In this study, the microbial composition of light-aroma-type Xiaoqu Baijiu at the saccharification and fermentation stages was investigated to explore microbial dynamics and their effects on aroma components using high-throughput sequencing and gas chromatography-flame ionization detection (GC-FID). Rhizopus, Pichia, Wickerhamomyces, Saccharomyces, Acinetobacter, Lactobacillus, and Weissella constituted the core microbes for Xiaoqu Baijiu production. Microbial succession during brewing could be divided into two phases: at the saccharification and early fermentation stages (F-0d to F-4d), Rhizopus and Acinetobacter were identified as the predominant microbes, accounting for 78.2-90.8% and 53.9-89.5% of the fungal and bacterial communities, respectively, whereas at the middle and late stages of fermentation (F-5d to F-14d), the abundance of Pichia, Wickerhamomyces, Saccharomyces, and Lactobacillus increased. Redundancy analysis (RDA) and Mantel tests indicated that the water, amino acid nitrogen, acid, and reducing sugar contents were significantly correlated with the fungal and bacterial communities in grains (p < 0.05). Pichia, Rhizopus, Saccharomyces, and Wickerhamomyces, especially Saccharomyces, were closely related to the contents of major alcohols, esters and aldehydes, and these microbes had an important functional role in the formation of Xiaoqu Baijiu flavor. This work provides insights into the microbial succession that occurs during brewing of light-aroma-type Xiaoqu Baijiu and the microbial contribution to flavor, which have potential for optimizing production and enhancing the flavor of Baijiu.

Cytoprotection against Oxidative Stress by Methylnissolin-3-O-ß-d-glucopyranoside from Astragalus membranaceus Mainly via the Activation of the Nrf2/HO-1 Pathway.

Astragalus membranaceus is a famous herb found among medicinal and food plants in East and Southeastern Asia. The Nrf2-ARE assay-guided separation of an extract from Jing liqueur led to the identification of a nontoxic Nrf2 activator, methylnissolin-3-O-ß-d-glucopyranoside (MNG, a component of A. membranaceus). Nrf2 activation by MNG has not been reported before. Using Western Blot, RT-qPCR and imaging, we investigated the cytoprotective effect of MNG against hydrogen peroxide-induced oxidative stress. MNG induced the expression of Nrf2, HO-1 and NQO1, accelerated the translocation of Nrf2 into nuclei, and enhanced the phosphorylation of AKT. The MNG-induced expression of Nrf2, HO-1, and NQO1 were abolished by Nrf2 siRNA, while the MNG-induced expression of Nrf2 and HO-1 was abated and the AKT phosphorylation was blocked by LY294002 (a PI3K inhibitor). MNG reduced intracellular ROS generation. However, the protection of MNG against the H2O2 insult was reversed by Nrf2 siRNA with decreased cell viability. The enhancement of Nrf2 and HO-1 by MNG upon H2O2 injury was reduced by LY294002. These data showed that MNG protected EA.hy926 cells against oxidative damage through the Nrf2/HO-1 and at least partially the PI3K/Akt pathways.

Effects of ultra-long fermentation time on the microbial community and flavor components of light-flavor Xiaoqu Baijiu based on fermentation tanks.

Microbial structure and succession of fermented grains play a significant role in Baijiu's flavor and quality. In this study, high-throughput sequencing (HTS) coupled with headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) were used to analyze the microbial community structures and flavor components in the fermented grains at the end of fermentation from different fermentation time of light-flavor Xiaoqu Baijiu. HTS results showed that Lactobacillus acetotolerans, Lactobacillus helveticus, Lactobacillus buchneri, Wickerhamomyces, Saccharomyces, and Condenascus were identified as the dominant microbes, but Lactobacillus (96.28%) exhibited obvious advantages at the end of ultra-long fermentation time (day 98). HS-SPME-GC-MS analysis revealed that esters and alcohols had the most abundance in fermented grains of day 98, containing high concentrations of ethyl acetate, diethyl succinate, phenylethyl alcohol, isoamyl alcohol, and n-propanol, which were related to the succession of Lactobacillus and yeast communities. Interestingly, the content of n-propanol in the ultra-long fermentation time samples (day 98) was 6 times of that in normal fermented grains (day 14), which may be caused by higher abundance of Lactobacillus in day 98 samples. Monte Carlo permutation test showed residual starch, acidity, and amino nitrogen (p < 0.05) were important factors affecting the microbial community. Together, these results shed light on the physicochemical changes, microbial dynamics, and key flavor components of fermented grains at the end of fermentation from different fermentation time and provide a strategy for further improvement of Baijiu quality.

Role of Alcohol Drinking in Alzheimer's Disease, Parkinson's Disease, and Amyotrophic Lateral Sclerosis.

Neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS), increase as the population ages around the world. Environmental factors also play an important role in most cases. Alcohol consumption exists extensively and it acts as one of the environmental factors that promotes these neurodegenerative diseases. The brain is a major target for the actions of alcohol, and heavy alcohol consumption has long been associated with brain damage. Chronic alcohol intake leads to elevated glutamate-induced excitotoxicity, oxidative stress and permanent neuronal damage associated with malnutrition. The relationship and contributing mechanisms of alcohol with these three diseases are different. Epidemiological studies have reported a reduction in the prevalence of Alzheimer's disease in individuals who drink low amounts of alcohol; low or moderate concentrations of ethanol protect against ß-amyloid (Aß) toxicity in hippocampal neurons; and excessive amounts of ethanol increase accumulation of Aß and Tau phosphorylation. Alcohol has been suggested to be either protective of, or not associated with, PD. However, experimental animal studies indicate that chronic heavy alcohol consumption may have dopamine neurotoxic effects through the induction of Cytochrome P450 2E1 (CYP2E1) and an increase in the amount of α-Synuclein (αSYN) relevant to PD. The findings on the association between alcohol consumption and ALS are inconsistent; a recent population-based study suggests that alcohol drinking seems to not influence the risk of developing ALS. Additional research is needed to clarify the potential etiological involvement of alcohol intake in causing or resulting in major neurodegenerative diseases, which will eventually lead to potential therapeutics against these alcoholic neurodegenerative diseases.

Hormesis of glyceollin I, an induced phytoalexin from soybean, on budding yeast chronological lifespan extension.

Glyceollin I, an induced phytoalexin isolated from soybean, has been reported to have various bioactivities, including anti-bacterial, anti-nematode, anti-fungal, anti-estrogenic and anti-cancer, anti-oxidant, anti-inflammatory, insulin sensitivity enhancing, and attenuation of vascular contractions. Here we show that glyceollin I has hormesis and extends yeast life span at low (nM) doses in a calorie restriction (CR)-dependent manner, while it reduces life span and inhibits yeast cell proliferation at higher (µM) doses. In contrast, the other two isomers (glyceollin II and III) cannot extend yeast life span and only show life span reduction and antiproliferation at higher doses. Our results in anti-aging activity indicate that glyceollin I might be a promising calorie restriction mimetic candidate, and the high content of glyceollins could improve the bioactivity of soybean as functional food ingredients.

The {332}<113> Twinning Behavior of a Ti-15Mo Medical Alloy during Cyclic Deformation and Its Effect on Microstructure and Performance.

In this study, the microstructural evolution of a Ti-15Mo medical alloy was investigated, when the in situ cyclic tensile strain had 2% amplitude and the tension-compression cyclic deformation had 1%, 2%, and 3% amplitude. The Vickers hardness and wear resistance of the alloy were also optimized due to the grain-refining effect after cyclic deformation and annealing. The twinning-induced plasticity (TWIP) was considered the main deformation mechanism of the Ti-15Mo alloy during the tensile-compressive cycle deformation with suitable strain amplitude. The {332}<113> twins and boundaries were the main contributors to the grain refinement. The optimal microstructure, hardness, and wear resistance were obtained in the alloy deformed by tension-compression cyclic strain with a 3% strain amplitude. The wear resistance of the annealed alloy in Hank's solution was excellent in contrast to the original Ti-15Mo alloy due to its reasonable microstructure and hardness. It is clear that abundant twins were formed and retained in the coarse grains of the original alloy after cyclic deformation and annealing, which provided the expected refined grains and performance.

Qingfei mixture modulates the immune responses in lung cancer through modulating mTOR signaling and gut microbiota-derived short-chain fatty acids.

Lung cancer ranks among the primary contributors to cancer-related fatalities on a global scale. Multiple research investigations have demonstrated that there exists a dysbiosis within the intestinal bacteria and short-chain fatty acids (SCFAs) is linked with immune responses in lung cancer. Qingfei mixture (QFM) has been widely used in treating lung cancer, yet the active ingredients and roles of the QFM on immune responses by targeting gut microbiota remain to be elucidated. The chemical constituents of QFM were qualitatively examined by UPLC/Q-TOF-MS. Additionally, we evaluated the therapeutic impact of the organic substance QFM on lung cancer, aiming to elucidate its mechanisms for improving the tumor-immune microenvironment. Herein, we constructed a Lewis lung carcinoma (LLC)-bearing mice model with QFM treatment to observe tumor growth and immune cell changes. Then, the feces were collected and a combinatory study using metagenomes, non-targeted metabonomics, and targeted metabonomics of SCFAs was performed. In vitro experiments have been conducted to estimate the roles of acetate and sodium propionate in CD8+ T cells. Furthermore, we treated tumor-bearing mice with QFM, QFM + MHY1485 (an mTOR activator), and QFM + an antibiotic mixture (ABX) to explore the potential therapeutic benefit of regulation of the tumor microenvironment. A total of 96 compounds were obtained from QFM by UPLC/Q-TOF-MS. Besides, the findings demonstrated that QFM exhibited significant efficacy against lung cancer, manifesting in reduced tumor growth and improved immune responses. In investigating its mechanisms, we integrated gut microbiota sequencing and fecal metabolomics, revealing that QFM effectively restored disruptions in gut microbiota and SCFAs in mice with lung cancer. QFM, acetate, or sodium propionate contributed to the up-regulation of IFN-γ, Gzms-B, perforin, IL-17, IL-6, IL-12, TNF-α expressions and decreased HDAC and IL-10 levels in vitro and in vivo. Moreover, MHY1485 and ABX weakened the effects of QFM on immunomodulation. Collectively, these results suggest that QFM may facilitate immune responses in the LLC-bearing mice via regulating the gut microbiota-derived SCFAs at least partially through targeting the mTOR signaling pathway.

Secondary metabolites in durian seeds: oligomeric proanthocyanidins.

Ethanolic extract of durian seeds was fractionated by reverse phase flash column chromatography and the fractions characterized by electrospray ionization mass spectroscopy. Among a few unknown compounds collected, oligomeric proanthocyanidins (OPCs) were found to be one of the main compounds. Based on this result, the OPCs were purified the first time from the durian seeds using standard procedures and gave a yield of 1.8 mg/g dry matter after fractionation by Sephadex LH-20 column. Structural analysis by ¹³C{¹H} NMR and ESI-MS spectra showed the presence of primarily B-type procyanidins with mainly epicatechin as the extension units, which was further verified by matrix assisted laser desorption/ionization-time of flight mass spectra (MALDI-TOF MS), which shows a distribution of dimers to decamers. In addition, hydroxylated peaks with molecular weight 16 units more than the poly-epicatechins represented significant peaks. We suggest this might be due to hydroxylation occurring under the MALDI-TOF MS conditions. Consistently, depolymerization with α-toluenethiol resulted in epicatechin thioether as the major product, but undetectable amount of gallocatechin or its α-toluenethiol derivatives. The oligomershave a mean degree of polymerization of 7.30.

Characterization of proanthocyanidins in stems of Polygonum multiflorum Thunb as strong starch hydrolase inhibitors.

Characterization of polyphenolic compounds in the stems of P. multiflorum was conducted using HPLC, high resolution LC-MS and LC-MSn. Proanthocyanidins in particular were isolated in 4.8% yield using solvent extraction followed by Sephadex LH-20 fractionation. HPLC analysis using a diol column revealed oligomers (from dimer to nonamer) as minor components, with (epi)catechin monomeric units predominating, and oligomers with higher degree of polymerization being dominant. Thiolysis treatment of the proanthocyanidins using mercaptoacetic acid produced thioether derivatives of (epi)catechin as the major product and a mean value of the degree of polymerization of 32.6 was estimated from the ratio of terminal and extension units of the (epi)catechin. The isolated proanthocyanidins were shown to strongly inhibit α-amylase with an acarbose equivalence (AE) value of 1,954.7 µmol AE/g and inhibit α-glucosidase with an AE value of 211.1 µmol AE/g.

Sulforaphene suppressed cell proliferation and promoted apoptosis of COV362 cells in endometrioid ovarian cancer.

Aim: N6-methyladenosine (m6A) RNA methylation exerts a regulatory effect on endometrioid ovarian cancer (EOC), but the specific m6A regulator genes in EOC remain to be explored. This study investigated that sulforaphene (Sul) is implicated in EOC development by regulating methyltransferase-like 3 (METTL3). Methods: The dysregulated m6A RNA methylation genes in EOC were determined by methylated RNA immunoprecipitation (MeRIP-seq) and RNA sequencing. The roles of METTL3 and/or Sul on viability, proliferative ability, cell cycle, and apoptosis of EOC cells were determined by MTT, colony formation, flow cytometry, and TUNEL staining assay, respectively. The expression of METTL3 and apoptosis-related proteins in EOC cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assays. Results: Five m6A RNA methylation regulators (METTL3, ELF3, IGF2BP2, FTO, and METTL14) were differentially expressed in EOC, among which METTL3 had the highest expression level. Silencing METTL3 reduced the clonal expansion and viability of EOC cells, and caused the cells to arrest in the G0/G1 phase. This also promoted apoptosis in the EOC cells and activated the FAS/FADD and mitochondrial apoptosis pathways. In contrast, overexpressing METTL3 had the opposite effect. Sul, in a dose-dependent manner, reduced the viability of EOC cells but promoted their apoptosis. Sul also increased the levels of IGF2BP2 and FAS, while decreasing the levels of KRT8 and METTL3. Furthermore, Sul was able to reverse the effects of METTL3 overexpression on EOC cells. Conclusions: Sul could suppress cell proliferation and promote apoptosis of EOC cells by inhibiting the METTL3 to activate the FAS/FADD and apoptosis-associated pathways.

Hangover headache and its behavioral changes in rats.

Objectives: The present study aims to establish and evaluate a rat model for hangover headaches caused by alcoholic drinks. Materials and Methods: Chronic migraine (CM) model rats were divided into 3 groups, and intragastrically administered alcoholic drinks (sample A, B, or C) to simulate hangover headache attacks. The withdrawal threshold for the hind paw/face and the thermal latency of hind paw withdrawal were detected after 24 hr. Serum was collected from the periorbital venous plexus of rats in each group, and enzymatic immunoassays were used to determine the serum levels of calcitonin gene-related peptide (CGRP), substance P (SP), and nitric oxide (NO). Results: Compared with the control group, the mechanical hind paw pain threshold was significantly lower in rats administered Samples A and B after 24 hr; however, no significant difference was observed across groups for the thermal pain threshold. The mechanical threshold for periorbital pain was only significantly reduced in rats administered Sample A. Immunoassays further indicated that serum levels of SP in the group administered Sample A were significantly higher than those in the control group; the serum levels of NO and CGRP were significantly higher in the group of rats receiving Sample B. Conclusion: We successfully developed an effective and safe rat model for investigating alcohol drink induced hangover headaches. This model could be used to investigate the mechanisms associated with hangover headaches for the development of novel and promising candidates for the future treatment or prophylaxis of hangover headaches.

Therapeutic strategies of small molecules in the microbiota-gut-brain axis for alcohol use disorder.

The microbiota-gut-brain axis (MGBA) is important in maintaining the structure and function of the central nervous system (CNS) and is regulated by the CNS environment and signals from the peripheral tissues. However, the mechanism and function of the MGBA in alcohol use disorder (AUD) are still not completely understood. In this review, we investigate the underlying mechanisms involved in the onset of AUD and/or associated neuronal deficits and create a foundation for better treatment (and prevention) strategies. We summarize recent reports focusing on the alteration of the MGBA in AUD. Importantly, we highlight the properties of small-molecule short-chain fatty acids (SCFAs), neurotransmitters, hormones, and peptides in the MGBA and discusses their usage as therapeutic agents against AUD.

Chemical Stability of a Chinese Herbal Spirit Using LC-MS-Based Metabolomics and Accelerated Tests.

As a prevalent medicinal liquor among Chinese people, a type of Chinese herbal spirit from Jing Brand Co., Ltd (CHS-J) is a newly developed health beverage with the health functions of anti-fatigue and immune enhancement. The researchers from the enterprise found that the contents of several components in CHS-J samples have been significantly decreasing during the stated storage period, as detected by the HPLC-UV method, which would make a great challenge for quality control of CHS-J. Furthermore, the chemical stability of CHS-J during the storage period is greatly challenged affected, especially in the environment of high temperature and light exposure. To systematically reveal the unstable components and promote the quality control of CHS-J, the chemical stability of CHS-J during the shelving storage period was characterized by the UPLC/Q-TOFMS-based metabolomics approach. First, the targeted and untargeted metabolomics approaches discovered the significantly changed components in CHS-J samples produced in different years. Furthermore, the accelerated tests of newly produced CHS samples and several authorized standards were conducted to validate the above results and elucidate the possible mechanisms underlying these chemical changes. Moreover, these chemical changes during the storage period had little influence on the anti-fatigue effect of CHS-J samples. These findings will offer new insight into the understanding of the chemical stability of CHS-J and will facilitate the quality control of CHS-J.

Simultaneous quantification of bioactive components in Chinese herbal spirits by ultra-high performance liquid chromatography coupled to triple-quadrupole mass spectrometry (UHPLC-QQQ-MS/MS).

BACKGROUND: The Chinese medicinal wine made from herbal medicines became prevalent among Chinese people. The Chinese herbal spirit is composed of several herbal extracts, and has the certain health functions, such as anti-fatigue and immune regulation. The quality evaluation of Chinese herbal spirit is greatly challenged by the enormous and complex components with great structural diversity and wide range of concentration distribution. METHODS: An ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) with multiple reaction monitoring (MRM) method was developed to simultaneously determine forty-three bioactive components in the Chinese herbal spirits produced by year 2014 and 2018. RESULTS: Quantitative results showed that 11 components, i.e.., puerarin (5), purpureaside C (7), daidzin (8), echinacoside (9), acteoside (15), epimedin B (22), epimedin C (23), icariin (24), eugenol (27), chikusetsusaponin iva (30) and Z-ligustilide (40), significantly decreased along with the increasing years of storage, while 5 compounds, i.e.., geniposidic acid (1), protocatechuic acid (2), crustecdysone (14), daidzein (18) and icariside I (35), were basically stable in all samples across the years. CONCUSION: The established method allowing to simultaneously determined 43 components with wide structural diversity and trace amounts will facilitate the quality control research of Chinese herbal spirits.

Comparison of Potent Odorants in Traditional and Modern Types of Chinese Xiaoqu Liquor (Baijiu) Based on Odor Activity Values and Multivariate Analyses.

Predominant odorants in modern and traditional types of Chinese xiaoqu liquor (Baijiu) were identified and compared by the combined use of gas chromatography-olfactometry, odor activity values (OAVs), and multivariate analyses. A total of 79 aroma compounds were identified in a typical modern type xiaoqu Baijiu (M) and a typical traditional type xiaoqu Baijiu (T), 42 of them had OAV > 1 in both M and T samples. The main differences between the two samples were obtained for the concentration of 23 aroma-active compounds. A total of 22 samples made by different brewing processes were analyzed to confirm the differences. Partial least squares discriminant analysis confirmed that 20 compounds could be used as potential markers for discrimination between modern type xiaoqu Baijiu and traditional type xiaoqu Baijiu. Their difference in content is between 1.5 and 17.9 times for modern type xiaoqu Baijiu and traditional type xiaoqu Baijiu. The results showed the aroma characteristics of modern and traditional type xiaoqu Baijiu clearly and comprehensively, which will provide guidance for modern Baijiu quality control and evaluation.

Quercetin in Tartary Buckwheat Induces Autophagy against Protein Aggregations.

Tartary buckwheat is used as an ingredient in flour and tea, as well as in traditional Chinese medicine for its antioxidant effects. Here, we found that an ethanol extract of tartary buckwheat (TBE) potently induced autophagy flux in HeLa cells by suppressing mTORC1 activity, as revealed by dephosphorylation of the mTORC1 substrates Ulk1, S6K, and 4EBP, as well as by the nuclear translocation of transcriptional factor EB. In addition to non-selective bulk autophagy, TBE also induced aggrephagy, which is defined as autophagy against aggregated proteins. Quercetin is a flavonol found at high levels in TBE. We showed that quercetin induced both non-selective bulk autophagy and aggrephagy. These effects were also observed in Huh-7 cells derived from hepatocytes. Thus, aggrephagy induction by TBE and quercetin may relieve alcoholic hepatitis, which is closely linked to the accumulation of protein aggregations called Mallory-Denk bodies.

Characterization of Key Aroma Compounds in Tartary Buckwheat (Fagopyrum tataricum Gaertn.) by Means of Sensory-Directed Flavor Analysis.

The key odorants of tartary buckwheat (TB) were researched by a sensory-directed flavor analysis approach for the first time. After the volatiles of TB were isolated by solvent-assisted flavor evaporation (SAFE), 49 aroma-active components with flavor dilution (FD) factors in the range of 1-2187 were identified using gas chromatography-olfactometry-mass spectrometry (GC-O-MS) combined with aroma extract dilution analysis (AEDA). Geranylacetone, phenethyl alcohol, and ß-damascone showed the highest FD factors of 2187. All 49 odorants were further quantitated by the internal standard curve method, and their odor activity values (OAVs) were obtained. The overall aroma of TB was successfully simulated (similarity > 98.16%) by mixing 16 odorants (OAV ≥ 1) with their natural concentrations. The omission tests revealed that geosmin, α-isomethylionone, α-methylionone, ß-ionone, linalool, ß-damascone, geranylacetone, guaiacol, ethyl hexanoate, geraniol, vanillin, tetrahydrolinalool, and 2,5-dimethyl-4-hydroxy-3-(2H)-furanone were the key odorants of TB. Chiral analysis showed that tetrahydrolinalool and linalool existed as racemics in the commercial TB. The relative content of R-enantiomers of α-isomethylionone and α-methylionone was slightly higher than that of their S-enantiomers. The odor thresholds of R- and S-enantiomer of tetrahydrolinalool were first detected as 0.029 and 3.8 µg/L in air, respectively.

Characterization of the key aroma compounds in Chinese JingJiu by quantitative measurements, aroma recombination, and omission experiment.

The unique flavor of and rich physiological activities exhibited by the Chinese JingJiu has made it become an essential part of the blended alcoholic beverage. In this study, the aromatic characteristics of Chinese JingJiu have been identified using sensory analysis, aroma extraction dilution analysis (AEDA), and quantitative analysis techniques. The odor activity values (OAVs) were also used to characterize the compound. A total of 136 aroma compounds were identified through the AEDA and gas chromatography-mass spectrometry (GC-MS) techniques. The flavor dilution (FD) factors were found to be in the range of 2-1024. Seventy aroma-active compounds with FD ≥ 8 were identified. Forty-three aroma-active compounds were identified using the molecular sensory science approach. Furthermore, 13 compounds were confirmed to be the key aroma-active compounds present in the Chinese JingJiu. The work provides a certain guiding effect on the regulation and optimization of the Chinese JingJiu production process.

A strategy for rapid discovery of traceable chemical markers in herbal products using MZmine 2 data processing toolbox: A case of Jing Liqueur.

Objective: The quality evaluation of herbal products remains a big challenge. Traceable markers are the core concept of the authentication of herbal products. However, the discovery of traceable markers is labor-intensive and time-consuming. The aim of this study is to develop a convenient approach to rapidly screen the traceable markers for herbal product authentication. Methods: Commercial Jing Liqueur and its 22 species of herbal ingredients were analyzed using HPLC-QTOF-MS and GC-MS to characterize nonvolatile and volatile chemicals. The acquired data were imported into MZmine 2 software for mass detection, chromatogram building, deconvolution and alignment. The aligned data were exported into a csv file and then traceable markers were selected using the built-in filter function in Excel. Finally, the traceable markers were identified by searching against online databases or publications, some of which were confirmed by reference standards. Results: A total of 288 chemical features transferred from herbal materials to Jing Liqueur product were rapidly screened out. Among them, 52 markers detected by HPLC-QTOF-MS were annotated, while nine volatile markers detected by GC-MS were annotated. Moreover, 30 of these markers were confirmed by comparing with reference standards. A chemical fingerprint consisting of traceable markers was finally generated to ensure the authentication and quality consistency of Jing Liqueur. Conclusion: A strategy for rapid discovery of traceable markers in herbal products using MZmine 2 software was developed.

A Traditional Chinese Medicine Plant Extract Prevents Alcohol-Induced Osteopenia.

Traditional Chinese medicine (TCM) has been practiced in the treatment of bone diseases and alcoholism. Chronic excessive alcohol use results in alcohol-induced bone diseases, including osteopenia and osteoporosis, which increases fracture risk, deficient bone repair, and osteonecrosis. This preclinical study investigated the therapeutic effects of TCM herbal extracts in animal models of chronic excessive alcohol consumption-induced osteopenia. TCM herbal extracts (Jing extracts) were prepared from nine Chinese herbal medicines, a combinative herbal formula for antifatigue and immune regulation, including Astragalus, Cistanche deserticola, Dioscorea polystachya, Lycium barbarum, Epimedium, Cinnamomum cassia, Syzygium aromaticum, Angelica sinensis, and Curculigo orchioides. In this study, Balb/c male mice were orally administrated alcohol (3.2 g/kg/day) with/without TCM herbal extracts (0.125 g/kg, 0.25 g/kg, or 0.5 g/kg) by gavage. Our results showed that after 50 days of oral administration, TCM herbal extracts prevented alcohol-induced osteopenia demonstrated by µ-CT bone morphological analysis in young adults and middle-aged/old Balb/c male mice. Biochemical analysis demonstrated that chronic alcohol consumption inhibits bone formation and has a neutral impact on bone resorption, suggesting that TCM herbal extracts (Jing extracts) mitigate the alcohol-induced abnormal bone metabolism in middle-aged/old male mice. Protocatechuic acid, a natural phenolic acid in Jing extracts, mitigates in vivo alcohol-induced decline of alkaline phosphatase (ALP) gene expression in the bone marrow of Balb/c male mice and in vitro ALP activity in pre-osteoblast MC3T3-E1 cells. Our study suggests that TCM herbal extracts prevent chronic excessive alcohol consumption-induced osteopenia in male mice, implying that traditional medicinal plants have the therapeutic potential of preventing alcohol-induced bone diseases.