Nonalcoholic fatty liver disease (NAFLD), as a common chronic liver disorder, is prevalent in the world. Recent evidence demonstrates that the "gut-liver axis" is related well to the progression of NAFLD, which regards gut microbiota and the intestinal barrier as two critical factors correlated with NAFLD. Diammonium glycyrrhizinate (DG), a compound of the natural bioactive pentacyclic triterpenoid glycoside, is the main component of licorice root extracts. The anti-inflammatory and liver protection effects of DG have already been reported, but to date, the mechanism has not been fully elucidated. In this research, we observed that DG reduced body weight, liver steatosis, as well as hepatic inflammation in NAFLD model mice induced by a high-fat diet. Illumina sequencing of the 16S rRNA revealed that DG intervention notably altered the composition of the gut microbiota in NAFLD mice. The richness of gut microbiota was significantly increased by DG. Specifically, DG reduced the Firmicutes-to- Bacteroidetes ratio and the endotoxin-producing bacteria such as Desulfovibrio and elevated the abundance of probiotics such as Proteobacteria and Lactobacillus. DG could augment the levels of short-chain fatty acid (SCFA)-producing bacteria such as Ruminococcaceae and Lachnospiraceae and promote SCFA production. In addition, DG supplementation dramatically alleviated the intestinal low-grade inflammation. Meanwhile, DG improved the expression of tight junction proteins, the goblet cell number, and mucin secretion and sequentially enhanced the function of intestinal barrier. Collectively, the prevention of NAFLD by DG might be mediated by modulating gut microbiota and restoring the intestinal barrier.

Obesity is a worldwide epidemic metabolic disease. Gut microbiota dysbiosis and bile acids (BAs) metabolism disorder are closely related to obesity. Farnesoid X-activated receptor (FXR), served as a link between gut microbiota and BAs, is involved in maintaining metabolic homeostasis and regulating glucose and lipid metabolism. We previously reported that diammonium glycyrrhizinate (DG) could alter gut microbiota and prevent non-alcoholic fatty liver disease. However, it remains ambiguous how DG affects the gut microbiota to regulate host metabolism. In this present study, 16S rRNA Illumina NovaSeq and metabolomic analysis revealed that DG treatment suppressed microbes associated with bile-salt hydrolase (BSH) activity, which, in turn, increased the levels of taurine-conjugated BAs accompanied by inhibition of ileal FXR-FGF15 signaling. As a result, several obesity-related metabolism were improved, like lower serum glucose and insulin levels, increased insulin sensitivity, few hepatic steatosis and resistance to weight gain. Additionally, decreased level of serum lipopolysaccharide was observed, which contributed to a strengthened intestinal barrier. The effect of DG on weight loss was slightly enhanced in the antibiotics-treated obese mice. Collectively, the efficacy of DG in the treatment of obesity might depend on gut microbiota-conjugated BAs-FXR axis. Hence, it will provide a potential novel approach for the treatment of obesity.

Background Clonorchiasis is caused by eating of raw or undercooked freshwater fish containing the larvae of Clonorchis sinensis ; the Kato-Katz method is widely applied in diagnosis. The improvement of repeated Kato-Katz smears from multiple stool samples has been well illuminated in many helminths other than C. sinensis . Methods A cross-sectional investigation was implemented to capture the epidemiology and risk factors of clonorchiasis among middle school students in Qiyang county, China. Students with complete data of six Kato-Katz thick smears from two stool samples were included in this analysis. Data on the habits of eating raw freshwater fish were also collected and compared. Results Altogether, 397 students had complete information of six smears, out of which 394 reported the information on eating habits. According to the ‘gold’ standard by six smears, 77 students (19.4%) were detected with C. sinensis . However, only 45 (11.3%) were detected using a single smear, with an underestimation of 41.6% compared to the ‘gold’ standard. However, the geometric mean of eggs per gram of feces in detected cases was 126.4 in a single smear, overestimated by 105.2% compared to 61.6 by the ‘gold’ standard. The linear relationship between prevalence and infection intensity of detected cases based on different smears was significantly negative. The habits of eating raw freshwater fish in the false negative cases was similar to those in the detected cases, but these two groups had significantly higher levels for habits of eating raw freshwater fish than negative individuals. Conclusions In low endemicity situations, underestimation of C. sinensis infection could not be avoided based on a limited number of Kato-Katz smears. Thus, repeated smears from at least two stool samples should be considered when an individual eats raw freshwater fish, drug efficacy is evaluated or elimination of C. sinensis is verified. Additionally, when logistics are insufficient for multiple samples to be taken for diagnosis for survey and surveillance in the areas or populations of low endemicity, prevalence accuracy needs to be corrected.

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