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.

We conducted a clinical trial in 26 healthy adult women to evaluate the effect of foods containing placental extract ingested for 12 consecutive weeks on 9 constitutional types in Traditional Chinese Medicine.The body constitution analysis set consisted of 23 subjects (excluding 3 dropouts). Twelve-week ingestion of placental extract-containing foods improved individual scores for “Gentleness” type, a healthy constitution, and “Qi-deficiency,” “Yang-deficiency,” “Yin-deficiency,” “Wet-heat” and “Qi-depression” types, which are all mibyou constitution.The safety analysis set consisted of 25 subjects (excluding 1 dropout). Only one subject experienced an adverse event (exacerbation of urticaria, an underlying disease). No other safety concerns arose.

With the cooperation of Kurobe City, we conducted a basic survey using the nine classification of constitution and an intervention survey using a constitutional guidance program. The basic survey was conducted by the Kurobe group (Kurobe River alluvial fan spring water drinker n=155, 60.6±10.4 years old) and the non-Kurobe group (normal tap water drinker n=99, 50.7±12.8 living in the Hokuriku region without Kurobe City). A total of three surveys were conducted using the constitution Questionnaire (CCMQ-J). As a result, the Gentleness type, which is usually called the healthy constitution, was clearly higher in the Kurobe group, with 44.6 % in the Kurobe group and 22.5 % in the non-Kurobe group. In the proportion of each biased constitution(Mibyo constitution), the Kurobe group had less Wet-heat type (4.5 % vs 11.8 %), Qi-depression type (5.4 % vs 10.7 %), Yin-deficiency type (7.1 % vs 12.3 %) , and Phlegm-wetness type (5.8 % vs 9.6 %) than the non-Kurobe group. After the basic survey was completed, the Kurobe group was randomly divided into two groups, an intervention group (n=65, 62.3±9.3 years old) and a non-intervention group (n=68, 62.7±9.8 years old), and compared with the non-Kurobe group (control group) (n=80, 51.9±13.6 years old). The survey design was a simple comparative study, and the primary endpoint was a comparative study of changes in constitutional scores before and after intervention. As a result, in the Yang-deficiency type, a significant difference was observed between the intervention group, the non-intervention group and the control group at the first time after the intervention (One-way ANOVA p=0.04). No change was seen in the second survey after the intervention. In the comparison of Phlegm-wetness type between the three groups, there was a tendency in the amount of change in the constitutional score (One-way ANOVA p=0.087). From the above, it was suggested that the constitutional survey can not only grasp the health and the pre-symptomatic state, but also improve the pre-symptomatic by combining the guidance methods according to the constitution.

Placental extract has been attracting attention as a functional food. In this review, I specifically described the effects of porcine placenta extract on beauty, menopausal symptom, Late-Onset Hypogonadism syndrome (LOH syndrome), shoulder stiffness, knee pain, cognition and osteoporosis. The excellent actions of porcine placental extract are considered to bring light to men as well as middle-aged women.