Acetaminophen (APAP) overdose is one of the major causes of acute liver failure. Severe liver inflammation and the production of oxidative stress occur due to toxic APAP metabolites and glutathione depletion. Growing evidence has proved that vitamin D (VD) exerts anti-inflammatory and antioxidative functions. Our objective was to explore the protective role of calcitriol (VD3) in acute APAP-induced liver injury. Methods: Adult male mice were randomized into three groups; control (n = 8), APAP (n = 8), and VD3 group (n = 8). All mice, except controls, received oral administration of APAP (400 mg/kg) and were sacrificed 24 h later. In the VD3 group, calcitriol (10 µg/kg) was injected intraperitoneally 24 h before and after exposure to APAP. Blood samples were collected to assess serum aminotransferase and inflammatory cytokines [tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6)]. Liver tissues were analyzed for hepatic glutathione (GSH), malondialdehyde (MDA), and histopathology. Results: APAP administration significantly increased serum aminotransferase, inflammatory cytokines, and induced cellular inflammation and necrosis. APAP also depleted hepatic GSH and elevated oxidative stress, as indicated by high MDA levels. In the APAP group, 25% of the mice (two out of eight) died, while no deaths occurred in the VD3 group. Treatment with calcitriol significantly reduced serum aminotransferase, TNF-α, and IL-6 levels in the VD3 group compared to the APAP group. Additionally, VD3 effectively restored GSH reserves, reduced lipid peroxidation, and attenuated hepatotoxicity. Conclusions: These findings demonstrate that VD3 prevents APAP-induced acute liver injury and reduces mortality in mice through its anti-inflammatory and antioxidative activity. Thus, VD3 might be a novel treatment strategy for APAP-induced hepatotoxicity.

Testosterone deficiency has been reported to accelerate nonalcoholic fatty liver disease (NAFLD). However, there are minimal data on the risk of NAFLD in transgender women and the treatment of NAFLD in this population. This study aimed to investigate the treatment effects and the mechanisms of action of genistein and sex hormones in orchiectomized (ORX) rats with nonalcoholic steatohepatitis (NASH) induced by a high fat high fructose diet (HFHF). Seven-week old male Sprague-Dawley rats were randomly divided into 7 groups (n = 6 each group); 1) control group, 2) ORX + standard diet group, 3) HFHF group, 4) ORX + HFHF group, 5) ORX + HFHF diet + testosterone group (50 mg/kg body weight (BW) once weekly), 6) ORX + HFHF diet + estradiol group (1.6 mg/kg BW daily), and 7) ORX + HFHF diet + genistein group (16 mg/kg BW daily). The duration of treatment was 6 weeks. Liver tissue was used for histological examination by hematoxylin and eosin staining and hepatic fat measurement by Oil Red O staining. Protein expression levels of histone deacetylase3 (HDAC3) and peroxisome proliferator-activated receptor delta (PPARδ) were analyzed by immunoblotting. Hepatic nuclear factor (NF)-ĸB expression was evaluated by immunohistochemistry. Rats in the ORX + HFHF group had the highest degree of hepatic steatosis, lobular inflammation, hepatocyte ballooning and the highest percentage of positive Oil Red O staining area among all groups. The expression of HDAC3 and PPARδ was downregulated, while NF-ĸB expression was upregulated in the ORX + HFHF group when compared with control and ORX + standard diet groups. Testosterone, estradiol and genistein treatment improved histological features of NASH together with the reversal of HDAC3, PPARδ and NF-ĸB protein expression comparing with the ORX + HFHF group. In summary, genistein and sex hormone treatment could alleviate NASH through the up-regulation of HDAC3 and PPARδ, and the suppression of NF-ĸB expression.

Nonalcoholic steatohepatitis (NASH) is a progressive form of nonalcoholic fatty liver disease (NAFLD) that is characterized by hepatic inflammation and steatosis. Currently, limited data exist regarding the risk of NASH in transgender women and the treatment options for this particular population. The use of testosterone supplementation is unfavorable for transgender women, and estrogen supplementation is linked to an increased risk of breast cancer; thus, an isoflavone derivative compound known as “genistein” could serve as a viable substitute for a hormone supplement in this context. The purpose of this study was to investigate the treatment effects and mechanisms of actions of genistein and sex hormones in orchidectomized (ORX) rats with nonalcoholic steatohepatitis induced via a high-fat high-fructose diet (HFHF) model. Male Sprague-Dawley rats (n = 42) were randomly assigned into seven groups; control, ORX + standard diet, HFHF, ORX + HFHF, ORX + HFHF diet + testosterone (50 mg/kg body weight (BW) once weekly), ORX + HFHF diet + estradiol (1.6 mg/kg BW daily), and ORX + HFHF diet + genistein (16 mg/kg BW daily). The duration of the study was 6 weeks. Some parts of liver tissue were used for histological examination by H&E staining. The determination of fat accumulation was performed using Oil Red O staining. SREBP1c and FAS gene expression were quantified using real-time PCR technique. The levels of all types of peroxisome proliferator-activated receptors (PPARs; α, δ, γ), proteins, and signal transducer and activator of transcription 1 (STAT1) signaling pathway were determined by both immunoblotting and immunohistochemistry. Rats in the ORX + HFHF group had the highest degree of hepatic steatosis, lobular inflammation, and hepatocyte ballooning, and showed higher levels of genes related to de novo lipogenesis, including SREBP1c and FAS. The expression of PPARγ and STAT1 were upregulated, while the expression of PPARα and PPARδ were downregulated in the ORX + HFHF group. Testosterone, estradiol and genistein treatments improved NASH histopathology together with the reversal of all types of PPAR protein expressions. Interestingly, genistein decreased the levels of STAT1 protein expression more than those of testosterone and estradiol treatment. Genistein and sex hormone treatment could ameliorate NASH through the upregulation of PPARα, and PPARδ, and the suppression of PPARγ and STAT1 expression.

Despite advances in drug development, treatment for non-alcoholic steatohepatitis (NASH)-related liver fibrosis remains limited, underscoring the need for more effective therapeutic strategies. Gardenia jasminoides exhibits promising anti-inflammatory and anti-fibrotic properties, suggesting its potential as a therapeutic candidate for NASH-related liver fibrosis. Accordingly, this study aimed to investigate the therapeutic potential of Gardenia jasminoides fruit extract (GJE) in a rat model of NASH with fibrosis induced by a high-fat, high-fructose (HFHF) diet, and to elucidate the underlying molecular mechanisms. Male Sprague-Dawley rats were randomly assigned to four groups (n = 6/group): 1) Control group, fed a standard diet; 2) NASH with fibrosis group (NF), fed a HFHF diet for 20 weeks; 3) Low-dose GJE group (NF + Low dose GJE), treated with 0.119 g/kg BW of GJE; and 4) High-dose GJE group (NF+High dose GJE), treated with 0.239 g/kg BW of GJE. GJE was administered via intragastric gavage once daily from weeks 13–20. Serum markers of liver injury (ALT and AST) were assessed. Liver histology was examined to evaluate NASH and fibrosis severity. Immunohistochemistry was performed to detect NF-kB p65 and Col1a1 expression and protein levels of MMP-12, α-SMA, IL-13 and TGF-β1 were quantified. Serum ALT and AST levels were significantly increased in the NF group, which also showed the highest NASH histopathology scores and collagen deposition. GJE treatment significantly reduced ALT and AST levels in both low- and high- dose groups. The low-dose GJE group showed marked improvement in NASH histopathology, while both treatment groups exhibited reduced hepatic collagen deposition. GJE treatment significantly suppressed NF-kB p65 and Col1a1 expression, along with downregulation of MMP-12, α-SMA, IL-13 and TGF-β1 protein levels. In conclusion, GJE effectively ameliorated HFHF diet-induced NASH with fibrosis by modulating inflammatory and fibrogenic pathways.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTb), remains one of the leading causes of death globally, primarily affecting the lungs. Recent technological advancements have improved TB detection, particularly through the application of deep learning (DL) techniques. This research involves a pathologist using microscopy to identify acid-fast bacilli (AFB), combined with a proposed method that incorporates DL for TB detection. The DL models’ accuracy and efficiency are assessed and compared. The integration of DL aims to assist pathologists by providing an initial analysis, potentially streamlining their workflow. The proposed method provides the accuracy of 94%, which is equivalent to the traditional method. In contrast, the workload of pathologists, who is the bottleneck of the diagnosis, is considerably reduced. This novel approach not only demonstrates the potential of DL in healthcare but also emphasizes the significance of advanced diagnostic methods in addressing global health challenges like TB, ultimately improving healthcare outcomes and reducing mortality rates.