Effects of dietary arsenic exposure on liver metabolism in mice.

Arsenic, a ubiquitous environmental toxicant with various forms and complex food matrix interactions, can reportedly exert differential effects on the liver compared to drinking water exposure. To examine its specific liver-related harms, we targeted the liver in C57BL/6 J mice (n=48, 8-week-old) fed with arsenic-contaminated food (30 mg/kg) for 60 days, mimicking the rice arsenic composition observed in real-world scenarios (iAsV: 7.3%, iAsIII: 72.7%, MMA: 1.0%, DMA: 19.0%). We then comprehensively evaluated liver histopathology, metabolic changes, and the potential role of the gut-liver axis using human hepatocellular carcinoma cells (HepG2) and microbiota/metabolite analyses. Rice arsenic exposure significantly altered hepatic lipid (fatty acids, glycerol lipids, phospholipids, sphingolipids) and metabolite (glutathione, thioneine, spermidine, inosine, indole-derivatives, etc.) profiles, disrupting 33 metabolic pathways (bile secretion, unsaturated fatty acid biosynthesis, glutathione metabolism, ferroptosis, etc.). Pathological examination revealed liver cell necrosis/apoptosis, further confirmed by ferroptosis induction in HepG2 cells. Gut microbiome analysis showed enrichment of pathogenic bacteria linked to liver diseases and depletion of beneficial strains. Fecal primary and secondary bile acids, short-chain fatty acids, and branched-chain amino acids were also elevated. Importantly, mediation analysis revealed significant correlations between gut microbiota, fecal metabolites, and liver metabolic alterations, suggesting fecal metabolites may mediate the impact of gut microbiota and liver metabolic disorders. Gut microbiota and its metabolites may play significant roles in arsenic-induced gut-liver injuries. Overall, our findings demonstrate that rice arsenic exposure triggers oxidative stress, disrupts liver metabolism, and induces ferroptosis.

Effects of Intestinal Microbiota on the Biological Transformation of Arsenic in Zebrafish: Contribution and Mechanism.

Both the gut microbiome and their host participate in arsenic (As) biotransformation, while their exact roles and mechanisms in vivo remain unclear and unquantified. In this study, as3mt-/- zebrafish were treated with tetracycline (TET, 100 mg/L) and arsenite (iAsIII) exposure for 30 days and treated with probiotic Lactobacillus rhamnosus GG (LGG, 1 × 108 cfu/g) and iAsIII exposure for 15 days, respectively. Structural equation modeling analysis revealed that the contribution rates of the intestinal microbiome to the total arsenic (tAs) and inorganic As (iAs) metabolism approached 44.0 and 18.4%, respectively. Compared with wild-type, in as3mt-/- zebrafish, microbial richness and structure were more significantly correlated with tAs and iAs, and more differential microbes and microbial metabolic pathways significantly correlated with arsenic metabolites (P < 0.05). LGG supplement influenced the microbial communities, significantly up-regulated the expressions of genes related to As biotransformation (gss and gst) in the liver, down-regulated the expressions of oxidative stress genes (sod1, sod2, and cat) in the intestine, and increased arsenobetaine concentration (P < 0.05). Therefore, gut microbiome promotes As transformation and relieves As accumulation, playing more active roles under iAs stress when the host lacks key arsenic detoxification enzymes. LGG can promote As biotransformation and relieve oxidative stress under As exposure.

The fecal arsenic excretion, tissue arsenic accumulation, and metabolomics analysis in sub-chronic arsenic-exposed mice after in situ arsenic-induced fecal microbiota transplantation.

Arsenic can be specifically enriched by rice, and the health hazards caused by high arsenic rice are gradually attracting attention. This study aimed to explore the potential of microbial detoxification via gut microbiome in the treatment of sub-chronic arsenic poisoning. We first exposed mice to high-dose arsenic feed (30 mg/kg, rice arsenic composition) for 60 days to promote arsenic-induced microbes in situ in the gastrointestinal tract, then transplanted their fecal microbiota (FMT) into another batch of healthy recipient mice, and dynamically monitored the microbial colonization by 16S rRNA sequencing and ITS sequencing. The results showed that in situ arsenic-induced fecal microbiome can stably colonized and interact with indigenous microbes in the recipient mice in two weeks, and established a more stable network of gut microbiome. Then, the recipient mice continued to receive high-dose arsenic exposure for 52 days. After above sub-chronic arsenic exposure, compared with the non-FMT group, fecal arsenic excretion, liver and plasma arsenic accumulation were significantly lower (P < 0.05), and that in kidney, hair, and thighbone present no significant differences. Metabolomics of feces- plasma-brain axis were also disturbed, some up-regulated metabolites in feces, plasma, and cerebral cortex may play positive roles for the host. Therefore, microbial detoxification has potential in the treatment of sub-chronic arsenic poisoning. However, gut flora is an extremely complex community with different microorganisms have different arsenic metabolizing abilities, and various microbial metabolites. Coupled with the matrix effects, these factors will have various effects on the efflux and accumulation of arsenic. The definite effects (detoxification or non-detoxification) could be not assured based on the current study, and more systematic and rigorous studies are needed in the future.

Microbial Diversity Analyses of Fertilized Thitarodes Eggs and Soil Provide New Clues About the Occurrence of Chinese Cordyceps.

Chinese cordyceps is a well-known fungus-larva complex with medicinal and economic importance. At present the occurrence of Chinese cordyceps has not been fully illuminated. In this study, the microbial diversities of fertilized Thitarodes eggs from sites A (high occurrence rates of Chinese cordyceps), B (low occurrence rates), and C (no Chinese cordyceps) were analyzed using 16S rRNA and ITS gene-sequencing technique. The previous sequencing data of soil from the same sites were conjointly analyzed. The results showed that bacterial communities among the eggs were significantly different. The bacterial diversity and evenness were much higher on site A. Wolbachia was overwhelmingly predominant in the eggs of sites B and C, while Spiroplasma showed preference on site A. The fungal between-group differences in the eggs were not as significant as that of bacteria. Purpureocillium in Cordyceps-related families showed preference on site A. Wolbachia, Spiroplasma, and Purpureocillium were inferred to be closely related to Chinese cordyceps occurrence. Intra-kingdom and inter-kingdom network analyses suggest that closer correlations of microbial communities (especially closer fungal positive correlations) in fertilized eggs might promote Chinese cordyceps occurrence. Besides, metabolic pathway analysis showed that in fertilized eggs or soil the number of bacterial metabolic pathways with significant differences in every comparison between two sites was greater than that of fungi. Collectively, this study provides novel information about the occurrence of Chinese cordyceps, contributing to the large-scale artificial cultivation of Chinese cordyceps.

Harmine is an effective therapeutic small molecule for the treatment of cardiac hypertrophy.

Harmine is a ß-carboline alkaloid isolated from Banisteria caapi and Peganum harmala L with various pharmacological activities, including antioxidant, anti-inflammatory, antitumor, anti-depressant, and anti-leishmanial capabilities. Nevertheless, the pharmacological effect of harmine on cardiomyocytes and heart muscle has not been reported. Here we found a protective effect of harmine on cardiac hypertrophy in spontaneously hypertensive rats in vivo. Further, harmine could inhibit the phenotypes of norepinephrine-induced hypertrophy in human embryonic stem cell-derived cardiomyocytes in vitro. It reduced the enlarged cell surface area, reversed the increased calcium handling and contractility, and downregulated expression of hypertrophy-related genes in norepinephrine-induced hypertrophy of human cardiomyocytes derived from embryonic stem cells. We further showed that one of the potential underlying mechanism by which harmine alleviates cardiac hypertrophy relied on inhibition of NF-κB phosphorylation and the stimulated inflammatory cytokines in pathological ventricular remodeling. Our data suggest that harmine is a promising therapeutic agent for cardiac hypertrophy independent of blood pressure modulation and could be a promising addition of current medications for cardiac hypertrophy.

Association Between Prospective Registration and Quality of Systematic Reviews in Type 2 Diabetes Mellitus: A Meta-epidemiological Study.

Background: We sought to investigate the methodological and reporting quality of published systematic reviews describing randomized controlled trials in type 2 diabetes mellitus and analyze their association with status of protocol registration. Methods: We searched the PubMed database and identified non-Cochrane systematic reviews, with or without meta-analysis, reporting on type 2 diabetes mellitus and published between 2005 and 2018. We then randomly selected 20% of these reviews in each year, and performed methodological and reporting quality assessment using the Assessment of Multiple Systematic Review 2 (AMSTAR-2) checklist and Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) statement. We also conducted regression analyses to explore the association between characteristics of systematic reviews and AMSTAR-2 or PRISMA scores. Results: A total of 238 systematic reviews, including 33 registered and 205 non-registered articles, met the inclusion criteria and were subsequently reviewed. Analysis indicated an increase in both registered rates and quality of systematic reviews in type 2 diabetes mellitus over the recent years. With regards to methodological and reporting quality, we found higher scores in registered, relative to non-registered reviews (AMSTAR-2 mean score: 18.0 vs. 14.5, P = 0.000; PRISMA mean score: 20.4 vs. 17.6, P = 0.000). AMSTAR-2 and PRISMA scores were associated with registration status, country of the first author, and statistical results, whereas the proportion of discussing publication bias and reporting funding sources were <40% for both registered and non-registered systematic reviews. Conclusions: Methodological and reporting quality of systematic reviews in type 2 diabetes mellitus indicates an improvement in the recent years. However, the overall quality remains low, necessitating further improvement. Future studies are expected to pay more attention to prospective registration, description of publication bias and reporting of funding sources.

Deep learning-based artificial intelligence model to assist thyroid nodule diagnosis and management: a multicentre diagnostic study.

BACKGROUND: Strategies for integrating artificial intelligence (AI) into thyroid nodule management require additional development and testing. We developed a deep-learning AI model (ThyNet) to differentiate between malignant tumours and benign thyroid nodules and aimed to investigate how ThyNet could help radiologists improve diagnostic performance and avoid unnecessary fine needle aspiration. METHODS: ThyNet was developed and trained on 18 049 images of 8339 patients (training set) from two hospitals (the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China, and Sun Yat-sen University Cancer Center, Guangzhou, China) and tested on 4305 images of 2775 patients (total test set) from seven hospitals (the First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China; the Sixth Affiliated Hospital of Sun Yat-sen University, Guangzhou, China; the Guangzhou Army General Hospital, Guangzhou, China; the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China; the First Affiliated Hospital of Sun Yat-sen University; Sun Yat-sen University Cancer Center; and the First Affiliated Hospital of Guangxi Medical University, Nanning, China) in three stages. All nodules in the training and total test set were pathologically confirmed. The diagnostic performance of ThyNet was first compared with 12 radiologists (test set A); a ThyNet-assisted strategy, in which ThyNet assisted diagnoses made by radiologists, was developed to improve diagnostic performance of radiologists using images (test set B); the ThyNet assisted strategy was then tested in a real-world clinical setting (using images and videos; test set C). In a simulated scenario, the number of unnecessary fine needle aspirations avoided by ThyNet-assisted strategy was calculated. FINDINGS: The area under the receiver operating characteristic curve (AUROC) for accurate diagnosis of ThyNet (0·922 [95% CI 0·910-0·934]) was significantly higher than that of the radiologists (0·839 [0·834-0·844]; p<0·0001). Furthermore, ThyNet-assisted strategy improved the pooled AUROC of the radiologists from 0·837 (0·832-0·842) when diagnosing without ThyNet to 0·875 (0·871-0·880; p<0·0001) with ThyNet for reviewing images, and from 0·862 (0·851-0·872) to 0·873 (0·863-0·883; p<0·0001) in the clinical test, which used images and videos. In the simulated scenario, the number of fine needle aspirations decreased from 61·9% to 35·2% using the ThyNet-assisted strategy, while missed malignancy decreased from 18·9% to 17·0%. INTERPRETATION: The ThyNet-assisted strategy can significantly improve the diagnostic performance of radiologists and help reduce unnecessary fine needle aspirations for thyroid nodules. FUNDING: National Natural Science Foundation of China and Guangzhou Science and Technology Project.

Overview of Clinical Trials on Type 2 Diabetes Mellitus: A Comprehensive Analysis of the ClinicalTrials.gov Database.

PURPOSE: A better understanding of the current features of type 2 diabetes mellitus (T2DM)-related clinical trials is important for improving designs of clinical trials and identifying neglected areas of research. It was hypothesized that the trial registration policy promoted the designs of T2DM-related trials over the years. Therefore, this study aimed to present a comprehensive overview of T2DM-related clinical trials registered in the ClinicalTrials.gov database. METHODS: T2DM-related clinical trials registered in the ClinicalTrials.gov database were searched and assessed the characteristics of the relevant trials. We searched PubMed and Google Scholar for the publication statuses of the primary completed trials. RESULTS: Overall, 5117 T2DM-related trials were identified for analysis. Of the interventional trials, 71.5% had a primary treatment purpose while only 8.9% were prevention or health service. There were more interventional trials registered prior to patient recruitment between 2012 and 2019 than between 2004 and 2011 (44.6% vs 19.9%, P<0.001). The period between 2012 and 2019 also had more trials that enrolled <100 participants (59.2% vs 50.9%), were single-center studies (60.7% vs 50.6%), had non-randomized allocations (11.3% vs 6.3%), were open-label (49.2% vs 45.6%), and had smaller sample sizes than the period between 2004 and 2011 (all P<0.001). The five-year cumulative publication rates after primary completion of the trials were <40%. CONCLUSION: Although the ClinicalTrials.gov database did not include all clinical trials, the trials registered in the ClinicalTrials.gov database still accounted for most of the clinical studies. Encouragingly, more interventional trials were registered prior to patient recruitment over the years. The majority of T2DM-related clinical trials focused on drug-related treatment, and trials regarding prevention in T2DM should be promoted. More attention should be paid to improve the publication and dissemination of clinical trials results.

Current landscape of type 1 diabetes mellitus-related interventional clinical trials registered on ClinicalTrials.gov: a cross-sectional study.

AIMS: A better understanding of the current features of type 1 diabetes mellitus (T1DM)-related interventional clinical trials is important for improving clinical trial designs and identifying neglected research areas. Therefore, this study aimed to comprehensively assess T1DM-related interventional clinical trials registered in the ClinicalTrials.gov database. METHODS: In this cross-sectional study, T1DM-related clinical trials registered in the ClinicalTrials.gov database were searched on July 1, 2020. The characteristics of the relevant trials were assessed. PubMed and Google Scholar were used to search for publication statuses of primary completed studies. RESULTS: Overall, 1,421 T1DM-related interventional clinical trials were identified for analysis. Of those trials, 509 (35.8%) involved children and 912 (64.2%) involved only adults. Overall, 63.2% of trials enrolled < 50 participants and 61.9% were registered after patient recruitment. Most trials were single-centered (66.0%). The proportions of trials with children were higher than those with only adults with respect to the primary purpose of health service or prevention (13.6% vs. 4.8%), intervention of device (30.8% vs. 23.9%), education or lifestyle (28.9% vs. 11.3%), and dietary supplement (5.7% vs. 2.5%) (all P < 0.01). Only 24.0% of trials had available results after primary completion. The 5-year cumulative publication rate after primary trial completion was < 40%. CONCLUSIONS: T1DM-related interventional clinical trials registered in ClinicalTrials.gov were dominated by small single-center studies. Most trials lacked the availability of results and their respective publications. Large multicenter interventional clinical trials on T1DM are needed, and more attention should be paid to improve the publication and dissemination of clinical trials results.

The Current Landscape of Clinical Studies Focusing on Thyroid Cancer: A Comprehensive Analysis of Study Characteristics and Their Publication Status.

Background: A better understanding of the current characteristics of clinical trials on thyroid cancer (TC) is important to improve trial designs and identify neglected areas of research. However, there is a lack of a thorough understanding of the clinical studies on TC. Therefore, this study aimed to present a comprehensive overview of clinical trials on TC based on the ClinicalTrials.gov database and evaluate their publication status. Methods: We searched for TC-related clinical studies registered in the ClinicalTrials.gov database before December 2018 by using the keyword "thyroid cancer" and assessed the characteristics of the included trials. We searched the publication status of primary completed studies in PubMed and Google Scholar. Results: A total of 450 studies were identified for analysis, including 333 (74.0%) interventional studies and 117 (26.0%) observational studies. Interventional studies about TC were commonly non-randomized (67.6%), single-arm (55.6%), single-center (76.3%), and early-phase (60.0%) trials. The major category for which studies were performed was for target drug-related therapy (53.6%). In addition, 57.0% of the primary completed interventional studies were published. The published studies were more commonly primary completed studies after 2010 and used randomization and were less commonly designed as single-arm studies and were conducted in the USA/Canada, compared to non-published studies (P < 0.05 for all). The median time from primary completion to publication was 46.5 months, and the time decreased to 36.5 months after 2010. Studies conducted in the USA/Canada [odds ratio (OR) = 9.43, P = 0.020] and multi-center studies (OR = 6.55, P = 0.021) significantly increased the potential of publication in high-impact journals. Conclusions: High-quality, randomized phase 3 trials regarding TC are still insufficient. Therefore, more efforts are needed to improve the treatment of poor prognostic TC and timely publication.