Integrated cell metabolomics and network pharmacology approach deciphers the anti-testosterone deficiency mechanisms of Bushen Zhuanggu Tang.

Testicular dysfunction is distinguished by a deficiency in testosterone levels, which can be attributed to the occurrence of oxidative stress injury in Leydig cells. The empirical prescription known as Bushen Zhuanggu Tang, developed by a highly experienced traditional Chinese medicine practitioner with six decades of clinical expertize, aligns with the traditional Chinese medicine principle of "kidney governing bone". Researchers have demonstrated that the administration of BSZGT can effectively enhance testosterone production. The objective of this study is to investigate the potential anti-testicular dysfunction effects of BSZGT and elucidate its underlying mechanism in an in vitro setting. Specifically, the impact of oxidative stress induced by H2O2 on the activity and testosterone levels of Leydig cells (TM3) was examined. Furthermore, the utilization of UPLC-QE-Qrbitrap-MS enabled the identification of the involvement of BSZGT in various metabolic pathways, including arginine biosynthesis, amino acyl-tRNA biosynthesis, Alanine, aspartate and glutamine metabolism, and Citrate Cycle, through the modulation of 25 distinct metabolites. Additionally, a network pharmacological analysis was conducted to investigate the pivotal protein targets associated with the therapeutic effects of BSZGT. The findings demonstrate the identification of six key proteins (CYP19A1, CYP1B1, ALOX5, ARG1, XDH, and MPO) that play a significant role in augmenting testicular function through their involvement in the ovarian steroid production pathway. In summary, our study presents a comprehensive research methodology that combines cell metabonomics and network pharmacology to enhance the discovery of new therapeutic agents for TD.

Exploring Asthma Mechanism of Belamcanda Chinensis by "Dose-effect Weighted Coefficient" Network Pharmacology and Experiment Validation.

BACKGROUND: Asthma is a chronic inflammatory disease of the airways that seriously endangers human health. Belamcanda chinensis (BC), a traditional Chinese medicine, has been used to counteract asthma as it has been shown to possess anti-inflammatory and regulatory immunity properties. OBJECTIVE: The study aimed to investigate the mechanisms of action of BC in the treatment of asthma; a "dose-effect weighted coefficient" network pharmacology method was established to predict potential active compounds. METHODS: Information on the components and content of BC was obtained by UPLC-QEOrbitrap- MS spectrometry. Based on BC content, oral bioavailability, and molecular docking binding energy, dose-effect weighting coefficients were constructed. With the degree greater than average as the index, a protein-protein interaction (PPI) database was used to obtain the core key targets for asthma under dose-effect weighting. GO function and KEGG pathway analyses of the core targets were performed using DAVID software. Finally, MTT and ELISA assays were used to assess the effects of active components on 16HBE cell proliferation. RESULTS: The experimental results using the 16HBE model demonstrated BC to have a potential protective effect on asthma. Network pharmacology showed SYK, AKT1, and ALOX5 to be the main key targets, and Fc epsilon RI as the promising signaling pathway. Eight components, such as tectoridin, mangiferin, luteolin, and isovitexin were the main active compounds, Finally, we analyzed the LPS-induced 16HBE proliferation of each active ingredient. Based on the activity verification study, all five predicted components promoted the proliferation of 16HBE cells. These five compounds can be used as potential quality markers for asthma. CONCLUSION: This study provides a virtual and practical method for the simple and rapid screening of active ingredients in natural products.

Study of the Mechanism of Astragali Radix in Treating Type 2 Diabetes Mellitus and Its Renal Protection Based on Enzyme Activity, Network Pharmacology, and Experimental Verification.

Astragali Radix (AR) is a common Chinese medicine and food. This article aims to reveal the active role of AR in treating Type 2 diabetes mellitus (T2DM) and its renal protective mechanism. The hypoglycemic active fraction was screened by α-glucosidase and identified by UPLC-QE-Orbitrap-MS spectrometry. The targets and KEGG pathway were determined through the application of network pharmacology methodology. Molecular docking and molecular dynamics simulation technology were used for virtual verification. Subsequently, a mouse model of T2DM was established, and the blood glucose and renal function indexes of the mice after administration were analyzed to further prove the pharmacodynamic effect and mechanism of AR in the treatment of T2DM. HA was determined as the best hypoglycemic active fraction by the α-glucosidase method, with a total of 23 compounds identified. The main active components, such as calycoside-7-O-ß-D-glucoside, methylnisoline, and formononetin, were revealed by network pharmacology. In addition, the core targets and the pathway have also been determined. Molecular docking and molecular dynamics simulation techniques have verified that components and targets can be well combined. In vivo studies have shown that AR can reduce blood sugar levels in model mice, enhance the anti-inflammatory and antioxidant activities of kidney tissue, and alleviate kidney damage in mice. And it also has regulatory effects on proteins such as RAGE, PI3K, and AKT. AR has a good therapeutic effect on T2DM and can repair disease-induced renal injury by regulating the RAGE/PI3K/Akt signaling pathway. This study provides ideas for the development of new drugs or dietary interventions for the treatment of T2DM.

Mechanism of action of Huangbaichen Sanwei formulation in treating T2DM based on network pharmacology and molecular docking.

Huangbaichen Sanwei formulation (HBCS) has been reported to have a good hypoglycemic effect, but its pharmacological mechanism of action remains unclear. We used network pharmacology and molecular docking to explore the potential mechanism of action of HBCS against type-2 diabetes mellitus (T2DM). Fifty-five active components from HBCS interfered with T2DM. Twenty-five core targets, such as AKT1, INS, INSR, MAPK1 were identified. Enrichment analyses showed that HBCS was involved mainly including insulin receptor signaling pathway, extracellular region, and insulin-like growth factor receptor binding and other biological processes; common targets had roles in treating T2DM by regulating diabetic cardiomyopathy and insulin resistance. Molecular docking verified that components combined with core targets. HBCS play a part in treating T2DM through multiple components and targets at the molecular level, which lays a theoretical foundation for research using HBCS to treat T2DM. The components, predicted targets, and T2DM targets of HBCS were searched through databases, and common targets were determined. Further screening of the core targets was conducted through the establishment of a protein -protein interaction network. The core targets were analyzed by Gene Ontology (GO) annotation utilizing the DAVID platform. And the enrichment of signaling pathways was explored by employing the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Cytoscape 3.9.1 was employed to construct a "TCM-components-core target-pathway" network. Autodock Vina was used to dock molecules to compare the binding activity of active molecules with targets.

Coupling coordination and spatio-temporal pattern evolution between ecological protection and high-quality development in the Yellow River Basin.

The Yellow River Basin is one of the most important economic development belt and ecological management regions in China, it is of vital importance to study the coupling coordination between ecological protection and high-quality development. However, the systematic research from the perspective of ecological-production-living is still lacking. Therefore, a comprehensive evaluation index system including 29 indicators is constructed from ecological, production and living dimension. To evaluate the high-quality development level and coupling coordination degree of 61 cities in the Yellow River Basin, a comprehensive measurement model and coupling coordination model are established using the entropy weight TOPSIS method. With the help of ArcGIS, the spatial characteristics of high-quality development level and coupling coordination are visually illustrated. The results showed that: (1) From 2011 to 2020, the high-quality development of 61cities in the Yellow River Basin showed an increasing trend, and the level of upper and lower reaches was higher than that of the middle reaches. (2) According to the high-quality development level of 61 cities, it was divided into three types: sustainable growth type with 44 cities, the fierce fluctuation type with 11 cities and the other 6 cities was stable type. (3) The coupling coordination degree of ecology, production and living system also showed an increasing trend, while the degree was not high. (4)About the year-on-year growth rate of coupling coordination degree for 61 cities in 2020 compared with 2011, there are 19 cities more than 30 %, and 23 cities between 20% and 30 %, 11cities was 10%-20 %, the other 8 cities was less than 10 %. (5) There is a significant spatial difference in the level of high-quality development in the Yellow River Basin, while coupling coordination degree does not significant in spatial layout. Therefore, the development of different regions should adjust measures to local conditions, give full play to their advantages, and make up for their shortcomings to promote the overall development of the city.

A transcriptomics and network pharmacology approach to elucidate the mechanism of action of geniposide on carbon tetrachloride-induced liver injury in rats.

Geniposide, the main active component of Fructus Gardeniae (FG), is known to confer protection against liver diseases. Herein we explored the hepatoprotective effects of geniposide and elucidated its molecular mechanism by transcriptome RNA-seq and network pharmacology. Liver injury was modeled by intraperitoneally injecting CCl4 (0.15% prepared with refined peanut oil) at a dose of 1.5 mL/kg thrice a week; from the second week, rats were administered geniposide (20 mg/kg or 40 mg/kg) by gavage for 6 weeks. Serum and liver samples were then collected to assess liver function indicators and inflammatory factors and to observe pathological changes in the liver. The Illumina HiSeq 4000 platform was used to obtain transcriptome data from the liver tissue of rats after geniposide administration. Core targets and pathways related to the liver protection mechanism of geniposide were further analyzed by integrating transcriptomics and network pharmacology. Differentially expressed genes (DEGs), core targets, and signaling pathways were identified by methods such as q-PCR, molecular docking, and Western blotting. We found that after geniposide administration, the levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and inflammatory factors decreased in the model group, and liver injury cells be effectively repaired. RNA-seq data analysis showed that compared to control group, the model group reversed 1,451 DEGs; further, compared to model group, geniposide reversed 511 DEGs. Eight key targets, including PIK3R1, ACOX3, and EGF, were found through further analyses. Geniposide was determined to mainly regulate the PPAR signaling pathway, apoptosis signaling pathway, and MAPK signaling pathway in liver tissues. To summarize, the protective and restorative effects of geniposide on rat liver may seem to be related to its efficacy in inhibiting the activation of inflammatory pathways, thereby reducing cell apoptosis. Our findings should serve as the basis for the development of functional foods or drugs to prevent and treat liver diseases.

UPLC-QE-Orbitrap-Based Cell Metabolomics and Network Pharmacology to Reveal the Mechanism of N-Benzylhexadecanamide Isolated from Maca (Lepidium meyenii Walp.) against Testicular Dysfunction.

Testicular dysfunction (TDF) is characterized by testosterone deficiency and is caused by oxidative stress injury in Leydig cells. A natural fatty amide named N-benzylhexadecanamide (NBH), derived from cruciferous maca, has been shown to promote testosterone production. Our study aims to reveal the anti-TDF effect of NBH and explore its potential mechanism in vitro. This study examined the effects of H2O2 on cell viability and testosterone levels in mouse Leydig cells (TM3) under oxidative stress. In addition, cell metabolomics analysis based on UPLC-Q-Exactive-MS/MS showed that NBH was mainly involved in arginine biosynthesis, aminoacyl-tRNA biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis, the TCA cycle and other metabolic pathways by affecting 23 differential metabolites, including arginine and phenylalanine. Furthermore, we also performed network pharmacological analysis to observe the key protein targets in NBH treatment. The results showed that its role was to up-regulate ALOX5, down-regulate CYP1A2, and play a role in promoting testicular activity by participating in the steroid hormone biosynthesis pathway. In summary, our study not only provides new insights into the biochemical mechanisms of natural compounds in the treatment of TDF, but also provides a research strategy that integrates cell metabolomics and network pharmacology in order to promote the screening of new drugs for the treatment of TDF.

Physical Activity Types, Physical Activity Levels and Risk of Diabetes in General Adults: The NHANES 2007-2018.

BACKGROUND: Recreational activities show benefits for diabetes prevention, but work-related activity and the total amount of individual physical activity is rarely discussed. PURPOSE: The purpose of this study was to evaluate the participation in five typical physical activities (vigorous work activity, vigorous recreational activities, moderate work activity, moderate recreational activities, and walk/bicycle for transportation), as well as the weekly distribution of total physical activity intensity, and to explore the relationships between physical activity types, physical activity levels, and risk of diabetes. STUDY DESIGN: Cross-sectional study. METHODS: The self-reported physical activity data on specific domains of physical activity were acquired from individuals in the 2007-2018 National Health and Nutrition Examination Survey (NHANES) using the Physical Activity Questionnaire (PAQ). Diabetes status was assessed by self-reported medical diagnosis or medication usage, or a fasting glucose concentration ≥ 126 mg/dL (fasting is defined as no caloric intake for at least 8 h) or HbA1c ≥ 6.5%. Weighted logistic regression was used to investigate the associations between physical activity types, physical activity levels, and risk of diabetes. RESULTS: Diabetes was less prevalent in people who participated in physical activity and the risk of diabetes reduced progressively as total physical activity levels increased. Younger adults (20-44 years) and males reported a higher proportion of high-intensity physical activity participation. CONCLUSIONS: Our findings highlight the importance of a physically active lifestyle for preventing diabetes. Distinct types of physical activity had different effects on the risk of diabetes. A greater total physical activity level was related to a substantial reduction in diabetes risk.

Sleep and risk of hypertension in general American adults: the National Health and Nutrition Examination Surveys (2015-2018).

OBJECTIVE: The purpose of research is to investigate the associations of sleep factors separately and jointly with risk of hypertension. METHODS: The National Health and Nutrition Examination Surveys (NHANES) is a nationally representative survey. Participants aged over 20 years with complete and credible data from the NHANES 2015-2016 and 2017-2018 waves were included. Hypertension was assessed based on self-report medical diagnoses, or antihypertensive medication use, or systolic blood pressure ≥140 mmHg and/or diastolic blood pressure ≥90 mmHg. Sleep information (sleep duration, trouble sleeping, daytime sleepiness, self-reported snoring and sleep-related breathing issue) was obtained from household interview. RESULTS: Of 7426 participants, the mean (standard deviation) age was 48.0 (17.3) years, 3845 (51.8%) were females. The prevalence of hypertension was 32.8%, and lower in those with 7-9 h sleep, no trouble sleeping, no excessive daytime sleepiness, no snoring or sleep apnea symptoms, decreased as the quantity of healthy sleep factors increased. The self-reported short sleep (odds ratio [OR]: 1.25, 95% confidence interval [CI]: 1.02-1.54, P  = 0.032), trouble sleeping (OR: 1.53, 95% CI: 1.20 to 1.95, P  = 0.001), excessive daytime sleepiness (OR: 1.17, 95% CI: 1.01-1.35, P  = 0.041) and sleep apnea symptoms (OR: 1.33, 95% CI: 1.10-1.61, P  = 0.004) were associated with 25%, 53%, 17% and 33% increased risk of hypertension, respectively. Participants with a poor sleep pattern was associated with higher hypertension risk (OR: 2.47, 95% CI: 1.90-3.22, P  < 0.001). CONCLUSION: Sleep behaviors were cross-sectionally associated with a considerably higher hypertension risk.

Screening of the Active Compounds against Neural Oxidative Damage from Ginseng Phloem Using UPLC-Q-Exactive-MS/MS Coupled with the Content-Effect Weighted Method.

The neuroprotective properties of ginsenosides have been found to reverse the neurological damage caused by oxidation in many neurodegenerative diseases. However, the distribution of ginsenosides in different tissues of the main root, which was regarded as the primary medicinal portion in clinical practice was different, the specific parts and specific components against neural oxidative damage were not clear. The present study aims to screen and determine the potential compounds in different parts of the main root in ginseng. Comparison of the protective effects in the main root, phloem and xylem of ginseng on hydrogen peroxide-induced cell death of SH-SY5Y neurons was investigated. UPLC-Q-Exactive-MS/MS was used to quickly and comprehensively characterize the chemical compositions of the active parts. Network pharmacology combined with a molecular docking approach was employed to virtually screen for disease-related targets and potential active compounds. By comparing the changes before and after Content-Effect weighting, the compounds with stronger anti-nerve oxidative damage activity were screened out more accurately. Finally, the activity of the selected monomer components was verified. The results suggested that the phloem of ginseng was the most effective part. There were 19 effective compounds and 14 core targets, and enriched signaling pathway and biological functions were predicted. After Content-Effect weighting, compounds Ginsenosides F1, Ginsenosides Rf, Ginsenosides Rg1 and Ginsenosides Rd were screened out as potential active compounds against neural oxidative damage. The activity verification study indicated that all four predicted ginsenosides were effective in protecting SH-SY5Y cells from oxidative injury. The four compounds can be further investigated as potential lead compounds for neurodegenerative diseases. This also provides a combined virtual and practical method for the simple and rapid screening of active ingredients in natural products.

Evolution of pathogenicity in obligate fungal pathogens and allied genera.

Obligate fungal pathogens (ascomycetes and basidiomycetes) and oomycetes are known to cause diseases in cereal crop plants. They feed on living cells and most of them have learned to bypass the host immune machinery. This paper discusses some of the factors that are associated with pathogenicity drawing examples from ascomycetes, basidiomycetes and oomycetes, with respect to their manifestation in crop plants. The comparisons have revealed a striking similarity in the three groups suggesting convergent pathways that have arisen from three lineages independently leading to an obligate lifestyle. This review has been written with the intent, that new information on adaptation strategies of biotrophs, modifications in pathogenicity strategies and population dynamics will improve current strategies for breeding with stable resistance.

Study the mechanism of peimisine derivatives on NF-κB inflammation pathway on mice with acute lung injury induced by lipopolysaccharide.

Peimisine is one of the alkaloids in Fritillariae ussuriensis Bulbus, which has anti-acute lung injury effect. In order to obtain compounds with superior bio-activity, 14 new derivatives were obtained from peimisine, and the better activity compounds were screened by MTT method. It was found that boc-leucine mono peimisine ester monoamide (compound G, 25 µg/ml) had increased cell survival rate and reduced the TNF-α, IL-1ß, IL-6, and iNOS levels in RAW 264.7 by lipopolysaccharide (LPS)-stimulated. In vivo, LPS (10 mg/kg) was given intraperitoneally to establish ALI model, and compound G (2.5 or 10 mg/kg) was injected into mice as the experimental group. The results showed that after the compound G (10 mg/kg) treatment, the Wet / Dry ratio of the lung was reduced, and the expression of TNF-α, IL-1ß, IL-6 and iNOS was inhibited. Meanwhile, compound G (10 mg/kg) could increase the content of IκB protein and reduce the content of p65 protein in lung tissue by Western blot analysis, which may play an anti-acute lung injury role by inhibiting the activity of NF-κB signaling pathway. In conclusion, compound G could attenuate LPS-induced ALI in mice and it may become a new approach to treat ALI.

Relationship between Sleep and Hypertension: Findings from the NHANES (2007-2014).

BACKGROUND: To evaluate the association of sleep factors (sleep duration, self-reported trouble sleeping, diagnosed sleep disorder) and combined sleep behaviors with the risk of hypertension. METHODS: We analyzed 12,166 adults aged 30-79 years who participated in the 2007-2014 National Health and Nutrition Examination Survey. Sleep duration, self-reported trouble sleeping and sleep disorders were collected using a standardized questionnaire. We included three sleep factors (sleep duration, self-reported trouble sleeping and sleep disorder) to generate an overall sleep score, ranging from 0 to 3. We then defined the sleep pattern as "healthy sleep pattern" (overall sleep score = 3), "intermediate sleep pattern" (overall sleep score = 2), and "poor sleep pattern" (0 ≤ overall sleep score ≤ 1) based on the overall sleep score. The definition of hypertension was based on self-reported antihypertensive medication use or biological measurement (systolic blood pressure ≥140 mm Hg and/or diastolic blood pressure ≥90 mm Hg). We used weighted logistic regression models to investigate the associations between sleep and hypertension. RESULTS: The overall prevalence of hypertension was 37.8%. A short sleep duration (OR = 1.20, 95% CI: 1.08 to 1.33, p = 0.001), self-reported trouble sleeping (OR = 1.45, 95% CI: 1.28 to 1.65, p < 0.001) and sleep disorder (OR = 1.33, 95% CI: 1.07 to 1.66, p = 0.012) were related to the risk of hypertension. Poor sleep patterns were closely correlated with the risk of hypertension (OR = 1.90, 95% CI: 1.62 to 2.24). CONCLUSIONS: Participants with poor sleep patterns were associated with an increased risk for hypertension.

Effects of medicine food Fructus Gardeniae on liver and kidney functions after oral administration to rats for 12 weeks.

Fructus Gardeniae (FG) is medicine food widely used for the treatment and prevention of various diseases. However, in recent years, research has suggested that high doses of FG can cause hepatotoxicity and nephrotoxicity. To assess this potential toxicity in more depth, this study investigated the effects of decocted FG and two of its bioactive constituents (geniposide and genipin) on liver and kidney function in rats. Rats were intragastrically administered FG (330 mg/kg body weight), geniposide (50 mg/kg body weight), or genipin (50 mg/kg body weight) for 12 weeks. Changes in body weight, liver and kidney indices, biochemical indices, and inflammatory factors were monitored. In addition, pathological sections were assessed and the expression of caspase-3, NF-κBp65, COX-2, and iNOS was detected by immunohistochemistry and Western blot. It was found that the levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine, and urea nitrogen increased following administration of FG, geniposide, and genipin. Furthermore, the activities of superoxide dismutase and reduced glutathione decreased following treatment, while malondialdehyde levels increased. Pathological and immunohistochemical evaluations further confirmed that FG and its constituents may cause damage to the liver and kidneys. The mechanism study revealed that the protein level of inflammatory pathway increased and further promoted apoptosis, suggesting that it should not be taken orally for extended periods of time. PRACTICAL APPLICATIONS: Chinese medicine and food safety have always been public health concerns. Fructus Gardeniae (FG) is a plant with a dual-purpose as it is used as both a medicine and food. Medicinally, it has the effects of heat-clearing and detoxification. However, its adverse effects and related mechanisms are not clear, and this has potential safety implications. In this study, rats were treated with FG for 12 weeks and found that the long-term administration of FG or high dosing can lead to damage to liver and kidney function. Therefore, close attention must be paid to the dosage of FG in order to achieve a therapeutic effect and avoid adverse reactions. Thus, this study lays a foundation for the safety evaluation and clinical application of FG.

Artemisinin protects DPSC from hypoxia and TNF-α mediated osteogenesis impairments through CA9 and Wnt signaling pathway.

Dental pulp stem cells (DPSCs) possess the ability of multi-lineage differentiation, and are excellent sources of tissue engineering and regenerative medicine. Oxygen concentration and inflammation are two critical environmental factors that affect the osteogenic differentiation of DPSCs. We aimed to study the role of the antimalarial drug artemisinin on the osteogenic differentiation of human DPSCs under the hypoxia and inflammation conditions. We demonstrated that hypoxia (5% O2) and inflammation (20 ng/mL TNF-α), alone or in combination, significantly diminished in vitro cell survival and increased apoptotic rates. Notably, hypoxia and TNF-α exerted accumulative effect in suppressing the osteogenic differentiation of DPSCs, as evidenced by reduced expression levels of osteogenesis-associated genes including ALP, RUNX2 and OCN in osteogenic condition, as well as reduced mineral nodules formation as indicated by alizarin red staining. Artemisinin at the dose of 40 µM markedly reversed the suppression in cell survival caused by hypoxia or inflammation, and reduced apoptotic rates and the expressions of pro-apoptotic proteins. Additionally, artemisinin restored osteogenic differentiation of DPSCs under the hypoxia or/and inflammation conditions. Moreover, the beneficial effect of artemisinin was dependent on upregulated expression of CA9 and CA9-mediated antioxidant responses, as CA9 knockdown abolished the protective role of artemisinin on DPSC osteogenesis. Furthermore, while hypoxia or/and inflammation significantly inactivated the Wnt/ß-catenin signaling in DPSCs, additional exposure to artemisinin re-activated this pathway to promote osteogenic differentiation of DPSCs. Our results provide novel insight on the link between artemisinin and DPSC osteogenesis, and suggest promising artemisinin-based strategies for better dentin/pulp tissue engineering.

Deubiquitylase OTUD6B Governs pVHL Stability in an Enzyme-Independent Manner and Suppresses Hepatocellular Carcinoma Metastasis.

Hypoxia inducible factors (HIFs) are the key transcription factors that allow cancer cells to survive hypoxia. HIF's stability is mainly controlled by von Hippel-Lindau (pVHL)-mediated ubiquitylation. Unlike sporadic clear-cell renal carcinomas, VHL mutation is rarely observed in hepatocellular carcinoma (HCC) and the regulatory mechanisms of pVHL-HIF signaling remain elusive. Here, it is shown that deubiquitylase ovarian tumor domain-containing 6B (OTUD6B) suppresses HCC metastasis through inhibiting the HIF activity. OTUD6B directly interacts with pVHL, decreases its ubiquitylation and proteasomal degradation to reduce HIF-1α accumulation in HCC cells under hypoxia. Surprisingly, OTUD6B limits the ubiquitylation of pVHL independent of its deubiquitylase activity. OTUD6B couples pVHL and elongin B/C to form more CBCVHL ligase complex, which protects pVHL from proteasomal degradation. Depletion of OTUD6B results in the dissociation of CBCVHL complex and the degradation of pVHL by Trp Asp repeat and suppressors of cytokine signaling box-containing protein 1 (WSB1). In human HCC tissues, the protein level of OTUD6B is positively correlated with pVHL, but negatively with HIF-1α and vascular endothelial growth factor. Low expression of OTUD6B predicts poor patient survival. Furthermore, OTUD6B gene is a direct transcriptional target of HIF-1α and upregulated upon hypoxia. These results indicate a previously unrecognized feedback loop consisting of OTUD6B, pVHL, and HIF-1α, and provide insights into the targeted hypoxic microenvironment for HCC therapy.

Dynamic changes of enzymes involved in sugar and organic acid level modification during blueberry fruit maturation.

In blueberry, sugars and organic acids determine fruit organoleptic quality and drastically change during fruit maturation. This study examined enzymes involved in the metabolism of sugars and organic acids during the three maturation phases (green, pink and blue). During maturation, an increase in sugar (mainly fructose and glucose) was associated with up-regulation of VcSPP (CUFF.32787.1), VcSPS (CUFF.14989.1), and VcINV (gene.g3367.t1.1, CUFF.8077.1 and CUFF.47310.2). A decrease in citrate was associated with VcACLY (CUFF.27347.1 and CUFF.28772.1) in the acetyl-CoA pathway and with VcGAD (CUFF.15663.1 and CUFF.13757.1) and VcGLT (CUFF.6416.1) in the GABA shunt. A decrease in malate was associated with VcMDH (CUFF.30072.1, CUFF.18332.1 and CUFF.24878.1) involved in malate biosynthesis, and with VcADH (gene.g1507.t1.1, CUFF.3210.1 and gene.g30667.t1.1) as well as VcPDC (CUFF.47532.1) involved in fermentation. Multi-isoforms of enzymes were divergent and differentially regulated, suggesting that they have specialized functions in these pathways. The information will contribute to the understanding of blueberry organoleptic quality.

Screening of the Hepatotoxic Components in Fructus Gardeniae and Their Effects on Rat Liver BRL-3A Cells.

Fructus Gardeniae (FG) is a common Chinese medicine and food. However, the toxicity of FG has drawn increasing concern, especially its hepatotoxicity. The purpose of this study was to screen the hepatotoxic components of FG and evaluate their effects on rat liver BRL-3A cells. The chemical composition of FG was determined by HPLC-ESI-MS. CCK-8 assay was used to evaluate the cytotoxicity of ten chemical components from FG, and then the toxic components with significant inhibitory activity were selected for further study. The results showed that geniposide, genipin, genipin-1-gentiobioside, gardenoside, and shanzhiside all suppress cells viability. Apoptosis assays further indicated that geniposide and its metabolite genipin are the main hepatotoxic components of FG. Pretreatment of cells with geniposide or genipin increased the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The activities of superoxide dismutase (SOD) and glutathione (GSH) were decreased, while the malondialdehyde (MDA) level was increased. The cell contents of tumor necrosis factor (TNF-α), interleukin-6 (IL-6), and nitric oxide (NO) were also increased. Molecular docking simulations were used to investigate the mechanism of FG-induced hepatotoxicity, revealing that geniposide and genipin bind strongly to the pro-inflammatory factor TNFR1 receptor of the NF-κB and MAPK signaling pathways. The obtained results strongly indicate that the hepatotoxicity of FG is caused by iridoids compounds. Genipin had the most significant hepatotoxic effect. These toxic substances destroy the cell antioxidant defense system, increasing inflammatory injury to the liver cells and leading to apoptosis and even necrosis. Thus, this study lays a foundation for toxicology research into FG and its rational application.

Screening of the Active Component Promoting Leydig Cell Proliferation from Lepidium meyenii Using HPLC-ESI-MS/MS Coupled with Multivariate Statistical Analysis.

Lepidium meyenii is now widely consumed as a functional food and medicinal product, which is known as an enhancer of reproductive health. However, the specific chemical composition and mechanism of action for improving sexual function are unclear. The present study aims at screening and determining the potential compounds, which promote mouse leydig cells (TM3) proliferation. The partial least squares analysis (PLS) was employed to reveal the correlation between common peaks of high performance liquid chromatography (HPLC) fingerprint of L. meyenii and the proliferation activity of TM3. The results suggested that three compounds had good activities on the proliferation of TM3 and promoting testosterone secretion, there were N-benzyl-hexadecanamide, N-benzyl-(9z,12z)-octadecadienamide and N-benzyl-(9z,12z,15z)-octadecatrienamide which might be the potential bioactive markers related to the enhancing sexual ability functions of L. meyenii. The first step in testosterone synthesis is the transport of cholesterol into the mitochondria, and the homeostasis of mitochondrial function is related to cyclophilin D (CypD). In order to expound how bioactive ingredients lead to promoting testosterone secretion, a molecular docking simulation was used for further illustration in the active sites and binding degree of the ligands on CypD. The results indicated there was a positive correlation between the binding energy absolute value and testosterone secretion activity. In addition, in this study it also provided the reference for a simple, quick method to screen the promoting leydig cell proliferation active components in traditional Chinese medicine (TCM).