BACKGROUND: Mitochondria, which harbor their own genome (mtDNA), have attracted attention due to the potential of mtDNA copy number (mtDNA-CN) as an indicator of mitochondrial dysfunction. Although mtDNA-CN has been proposed as a simple and accessible biomarker for metabolic disorders such as metabolic dysfunction-associated steatotic liver disease, the underlying mechanisms and the causal relationship remain insufficiently elucidated. In this investigation, we combined longitudinal epidemiological data, animal studies, and in vitro assays to elucidate the potential causal relationship between reduced mtDNA-CN and the development of steatotic liver disease (SLD). METHODS: We conducted a longitudinal study using data from a health examination cohort initiated in 1981 in Yakumo, Hokkaido, Japan. Data from examinations performed in 2015 and 2022 were analyzed, focusing on 76 subjects without SLD at baseline (2015) to assess the association between baseline mtDNA-CN and subsequent risk of SLD development. In addition, 28-day-old SD rats were fed ad libitum on a 45% high-fat diet and dissected at 2 and 8 weeks of age. Blood and liver mtDNA-CN were measured and compared at each feeding period. Additionally, in vitro experiments were performed using HepG2 cells treated with mitochondrial function inhibitors to induce mtDNA-CN depletion and to examine its impact on intracellular lipid accumulation. RESULTS: Epidemiological analysis showed that the subjects with low mtDNA-CN had a significantly higher odds ratio for developing SLD compared to high (odds ratio [95% confidence interval]: 4.93 [1.08-22.50]). Analysis of the animal model showed that 8 weeks of high-fat diet led to the development of fatty liver and a significant decrease in mtDNA-CN. A further 2 weeks of high-fat diet consumption resulted in a significant decrease in hepatic mtDNA-CN, despite the absence of fatty liver development, and a similar trend was observed for blood. Complementary in vitro experiments revealed that pharmacologically induced mitochondrial dysfunction led to a significant reduction in mtDNA-CN and was associated with increases in intracellular lipid accumulation in HepG2 cells. CONCLUSIONS Our findings suggest that reduced mtDNA-CN may contribute causally to SLD development and could serve as a convenient, noninvasive biomarker for early detection and risk assessment.
Animals ; DNA, Mitochondrial/genetics* ; Humans ; Male ; DNA Copy Number Variations ; Female ; Fatty Liver/blood* ; Rats ; Middle Aged ; Longitudinal Studies ; Rats, Sprague-Dawley ; Adult ; Japan/epidemiology* ; Aged ; Biomarkers/blood* ; Hep G2 Cells ; Diet, High-Fat/adverse effects*

Objective: A study was conducted to clarify the anti-influenza virus effect of agricultural, forest and fishery products in Ishikawa Prefecture. Method: The samples were prepared by both hot-water and ethanol extraction. The Madin-Darby canine kidney (MDCK) cells infected with PR8 strain were cultured in Dulbecco's Modified Eagle Medium（DMEM） containing extract for 24 hours and measured virus titer of supernatants by focus-forming reduction assay. Time-of-addition assay was performed to determine the inhibition stage in virus proliferation by the extract. The active ingredients were purified using a solid-phase extraction column (silica gel base C18). Results: We revealed that many products have anti-influenza virus activity. Especially, four products ( Matteuccia struthiopteris, Colocasia esculenta, Sagittaria trifolia, and Eleucine coracana) show strong activity and inhibit several stages in virus proliferation. Conclusion: We performed screening to 46 agricultural, forest and fishery products and found the four products ( Matteuccia struthiopteris, Colocasia esculenta, Sagittaria trifolia, and Eleusine coracana) inhibit virus proliferation strongly. In addition, the isolation and identification of the active ingredients contained in these remain as a future study.

Objective: We investigated the effects of Coix-seed Reactive Derivatives (CRD) on fibroblast proliferation and collagen production. Methods: Various concentrations of CRD (0.00001%, 0.0001%, 0.001%, 0.01%, 0.1%, 1%) were added to human fibroblasts, and changes in cell count and extracellular collagen production were measured. Results: Fibroblasts proliferated by the addition of CRD, peaked at about 1.7 times when the CRD concentration was 0.0001%, and then decreased as the addition concentration increased. On the other hand, the collagen concentration in the extracellular matrix of fibroblasts increased as the CRD addition concentration increased, and the collagen concentration increased sharply at 0.1% of CRD concentration or more. Conclusion: It was suggested that CRD has a function of increasing the proliferation of fibroblasts and collagen production.

We evaluated the long-term safety of eight adult volunteers (male 7, female 1: average age 47.4 years) ingesting powdered foods containing Coix-seed Reactive Derivatives (CRD/2.2g/day or 4.4g/day) for 1 year 8 months to 5 years (average 2 years 9 months). Body weight, vital signs (blood pressure, pulse rate), peripheral blood test, blood biochemical test, etc. were observed. We also investigated the presence or absence of side effects. The results showed no notable changes in vital signs or blood test results. From the above, it was speculated that long-term intake of CRD would not pose a safety problem. We plan to continue the study by increasing the number of observation cases in the future.

Huaier granule, which is clinically used in Japan as a health food, is originally a traditional Chinese medicine for immune dysfunction, such as cancer, in China. Huaier guanule emerged with abundant evidence, and is being prescribed to hepatocellular carcinoma, breast cancer, lung cancer, gastric cancer, et al. as indications in China. This review focused on the clinical evidence of Huaier guanule associated to breast cancer.

Melinjo (Gnetum gnemon L.) is distributed in Southeast Asia and its fruits, seeds and leafs are commonly eaten in Indonesia. It was found that melinjo seed extract contains some resveratrol dimers (gnetin C, gnemonoside A and gnemonoside D). Melinjo seed extract and gnetin C have been reported to show beneficial effects on several diseases, such as hyperuricemia, atherosclerosis, fatty liver, diabetes, dementia, cancer, skin aging and periodontitis etc．Melinjo seed extract is one of the desired functional food materials in Japan.

To clarify the functions of Coix-seed Reactive Derivatives(CRD), we analyzed 13 kinds of phenolic compounds and coixol(an alkaloid) in CRD powder by using Liquid Chromatography/Mass Spectrometry(LC-MS).The following are the results: p -Coumaric acid 275.0 (μg/g), p -Hydroxybenzaldehyde 50.2 (μg/g), Caffeic acid 36.9 (μg/g), Protocatechuic acid 31.9 (μg/g), Ferulic acid 27.1 (μg/g), Quercetin 16.2 (μg/g), Syringaldehyde 12.1 (μg/g), Chlorogenic acid 11.3 (μg/g), Naringenin 11.1 (μg/g), Vanillin 9.2 (μg/g), trans -Coniferylaldehyde 9.1 (μg/g), Tricin 0.7 (μg/g), Luteolin 0.5 (μg/g) and Coixol(6-Methoxy-2-benzoxazolinone) 5.5 (μg/g). CRD is composed of many substances other than these phenoic compounds. Therefore, further studies are needed to understand the various clinical features associated with CRD intake.

We evaluated the effect of Coix-seed Reactive Derivatives (CRD) on cold sensitivity of female hands and feet. Ninety females were assigned randomly to 3 groups and CRD(2g/day, 4g/day or placebo) was administrated for 8weeks. Significant improvement of cold sensitivity was observed in both the 2g/day and 4g/day groups. It was speculated that a naringenin and a quercetin as active ingredients of adlay hull may play a critical role in ameliorating effect on blood flow.

Background: Adlay (Coix lacryma-jobi L. var. ma-yuen Stapf) has been used both in traditional Chinese medicine and as a nourishing food based on its unique biological effects and highly nutritional values. In the present study, we investigated the anti-tumor effect of a hot-water adlay extract in sarcoma mice model. Materials and Methods: The hot water extract of whole adlay was orally administered to mice for one week, after which Sarcoma-180 cells (1×106) were subcutaneously implanted into the abdomen. Thereafter, the tumor growth was monitored and mouse survival was examined. Results: Tumor weights measured at 18 days were significantly lower in mice treated with extract (100 and 300 mg/kg/day) than those in control group (p<0.01). Moreover, mice treated with extract (100 mg/kg/day) showed apparently longer survival than control group evaluated until 32 days (p<0.05). Conclusion: These findings indicate that hot water adlay extract appears to have some anti-tumor effects in vivo insarcoma cells.