Siraitia grosvenorii is the fruit of a cucurbitaceous vine endemic to China. Its extract has been used as a sweetener and exhibits various anti-inflammatory and anticarcinogenic effects mediated via its antioxidant properties. In the present study, we aimed to clarify the preventive or ameliorative effects of S. grosvenorii extract (SGE) on nonalcoholic steatohepatitis-like lesions induced in male Hsd: Sprague Dawley rats fed a choline-deficient, methionine-lowered, l-amino acid-defined diet for 13 weeks. This diet increased hepatotoxicity parameters and upregulated the expression of inflammation- and fibrosis-related genes in the liver, resulting in the progression of hepatic lesions, oxidative stress, hepatocellular apoptosis, and fibrosis. Furthermore, this diet upregulated the expression of phosphorylated nuclear factor-κB (NF-κB) and CD44. SGE administration inhibited these lesions, similar to CD44, a factor that controls hepatic inflammation and fibrosis. These results revealed that SGE impacts the disease stage via antioxidative effects and regulation of CD44 expression. SGE was found to be useful for preventing and treating steatohepatitis.

Laser resurfacing may be accompanied by unwanted side effects. The micro coring technology, designed to remove small skin columns, was developed to avoid the thermal injury associated with lasers. However, very limited data are available on its pre-clinical efficacy and safety. The novel robotic, fractional micro-coring device, Aime^TM, was tested on four pigs, each treated in 12 sites, at 6 time-points, over the course of 28 days. Macroscopic and microscopic evaluation was performed at each of the 6 time-points during the 28-day follow-up. Macroscopically, treatment resulted in erythema and mild edema that quickly resolved. Microscopically, there was progressive re-coverage of the tested sites with complete, well differentiated, newly formed epidermis, associated with efficient elimination of the underlying excised dermis, which was replaced by maturing fibroplasia. Some of the sites demonstrated complete healing already after 7 days. No significant adverse events were noted with the use of the device. The use of the micro-coring device Aime^TM in a porcine model for skin fractional micro-excision and resurfacing was effective and safe. The comprehensive gradual healing process shown in this study with detailed histopathological images can also serve as a basis for future pre-clinical studies of fractional ablative devices.

Recently, with the development of computer vision using artificial intelligence (AI), clinical research on diagnosis and prediction using medical image data has increased. In this study, we applied AI methods to analyze hepatic fibrosis in mice to determine whether an AI algorithm can be used to analyze lesions. Whole slide image (WSI) Sirius Red staining was used to examine hepatic fibrosis. The Xception network, an AI algorithm, was used to train normal and fibrotic lesion identification. We compared the results from two analyses, that is, pathologists’ grades and researchers’ annotations, to observe whether the automated algorithm can support toxicological pathologists efficiently as a new apparatus. The accuracies of the trained model computed from the training and validation datasets were greater than 99%, and that obtained by testing the model was 100%. In the comparison between analyses, all analyses showed significant differences in the results for each group. Furthermore, both normalized fibrosis grades inferred from the trained model annotated the fibrosis area, and the grades assigned by the pathologists showed significant correlations. Notably, the deep learning algorithm derived the highest correlation with the pathologists’ average grade. Owing to the correlation outcomes, we conclude that the trained model might produce results comparable to those of the pathologists’ grading of the Sirius Red-stained WSI fibrosis. This study illustrates that the deep learning algorithm can potentially be used for analyzing fibrotic lesions in combination with Sirius Red-stained WSIs as a second opinion tool in non-clinical research.

Our previous 4-week repeated dose toxicity study showed that wood preservative chromated copper arsenate (CCA) induced hepatocellular hypertrophy accompanied by biochemical hepatic dysfunction and an increase in oxidative stress marker, 8-hydroxydeoxyguanosine, in female rats. To further explore the molecular mechanisms of CCA hepatotoxicity, we analyzed 10%-buffered formalin-fixed liver samples from female rats for cell proliferation, apoptosis, and protein glutathionylation and conducted microarray analysis on frozen liver samples from female rats treated with 0 or 80 mg/kg/day of CCA. Chemical analysis revealed that dimethylated arsenical was the major metabolite in liver tissues of male and female rats. CCA increase labeling indices of proliferating cell nuclear antigen and decrease terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling accompanied with increased expression of protein glutathionylation, indicating a decrease in glutathione (GSH) in hepatocytes of female rats. Microarray analysis revealed that CCA altered gene expression of antioxidants, glutathione-S-transferase (GST), heat shock proteins and ubiquitin-proteasome pathway, cell proliferation, apoptosis, DNA methylation, cytochrome P450, and glucose and lipid metabolism in female rats. Increased expression of GSTs, including Gsta2, Gsta3, Mgst1, and Cdkn1b (p27), and decreased expression of the antioxidant Mt1, and DNA methylation Dnmt1, Dnmt3a, and Ctcf were confirmed in the liver of female rats in a dose-dependent manner. Methylation status of the promoter region of the Mt1 was not evidently changed between control and treatment groups. The results suggested that CCA decreased GSH and altered the expression of several genes, including antioxidants, GST, and DNA methylation, followed by impaired cell proliferation in the liver of female rats.

Spontaneous cartilaginous metaplasia of the sclera has not been reported in rabbits. Herein, we report two cases of spontaneous cartilaginous metaplasia in the sclera of Japanese White (JW) rabbits. Case 1 was noted in a 14-week-old male Kbs:JW rabbit that received a single ocular instillation of 20% isoproterenol (IP) a day before necropsy, and showed no abnormalities in clinical signs, ophthalmological assessments, and necropsy. Case 2 was noted in a 38-week-old male Kbs:JW rabbit that was housed under light-emitting diode (LED) lighting for 26 weeks and showed no effects of LED on clinical signs, ophthalmological assessments, and necropsy. Histological sections of the eyes of both animals were prepared and stained with hematoxylin and eosin (H&E) and Alcian blue, and immunohistochemical staining for vimentin was performed. The H&E-stained specimens showed focal hyaline cartilage-like tissues distributed between the scleral fibers at the posterior pole in both cases. The surrounding scleral fibers were compressed and/or partially destroyed by the cartilage-like tissue. The cartilage-like matrix was stained blue by Alcian blue, and immunohistochemistry showed that chondrocyte-like cells were positive for vimentin. Based on these findings, we diagnosed cartilaginous metaplasia in the sclera of Kbs:JW rabbits. The lesion was farther from the IP administration site in Case 1 and was not accompanied by other ophthalmological or histopathological abnormalities in either of the cases. This implies that the lesions occurred spontaneously owing to the abnormal differentiation of neural crest-derived cells.