Hepatocellular carcinoma has high fatality and poor prognosis. For curing hepatocellular carcinoma, the demand for effective therapeutic reagents with low toxicity is urgent. Herein, we investigated plasma-activated medium, an emerging reagent obtained via irradiation of cell-free medium with cold atmos­pheric plasma. Plasma-activated medium exerts inhibitory effect on many types of tumor cells with little toxicity to non-cancerous cells. In present study, we verified the tumor-specific inhibition of plasma-activated medium on hepatocellular carcinoma cell lines. Under the effect of plasma-activated medium, oxidative stress, mitochondrial dysfunction, and loss of intracellular NAD⁺ and ATP ‍were detected inside cells, suggesting an energy depletion. Through investigating the salvage pathway which synthesizes NAD⁺ and maintains the respiratory chain in hepatocellular carcinoma, we found that the energy failure was resulted by the ‍blockage of the salvage pathway. Moreover, nicotinamide phosphoribosyltransferase, the rate-limiting enzyme in the salvage pathway, was determined as an important target to be inactivated by the effect of plasma-activated medium. Additionally, the blockage of the salvage pathway activates AMPKα and suppresses mTOR pathway, which reinforces the cell growth inhibition. Overall, our findings demonstrated that the disruption of functions of nicotinamide phosphoribosyltransferase and the salvage pathway contribute to the tumor-specific cytotoxicity of plasma-activated medium.

Mitophagy plays a vital role in carcinogenesis and tumor progres­sion. However, the research on the mechanism of mitophagy in esopha­geal cancer metastasis is limited. This study explored the regulatory mechanism of RECQL4 in mitophagy and affects esopha­geal cancer metastasis. The RECQL4 expression in esopha­geal cancer tissues and cells was examined by bioinformatics and qRT-PCR. Bioinformatics analysis was used to determine the upstream regulatory factor of RECQL4 and CREB1. Their binding relationship was evaluated by dual luciferase and Chromatin Immunoprecipitation assays. The effects of RECQL4 on esophageal cancer cells viability, metastasis, and mitophagy were examined using CCK-8, Transwell, immunofluorescence, and Western blot assays. The expression of RECQL4 was up-regulated in esophageal cancer tissues and cells. Overexpression of RECQL4 promoted the cells viability, invasion, migration, and epithelial-mesenchymal transition by inhibiting mitophagy. Bioinformatics analysis revealed a positive correlation between RECQL4 and CREB1, their binding relationship was validatied by dual luciferase and ChIP assays. CREB1 knockdown promoted mitophagy and prevented the metastasis of cancer cells, which could be countered by overexpressing RECQL4. In conclusion, CREB1 was found to transcriptionally activate RECQL4 to inhibit mitophagy, thereby promoting esophageal cancer metastasis. Targeting mitophagy could be an effective therapeutic approach for esophageal cancer.

Neutrophils play an important role in innate immunity and produce reactive oxygen species, but they can also cause inflam­mation and oxidative stress that can damage their own tissues. We have developed neutrophil activity evaluation systems that simultaneously monitors superoxide radicals and hypochlorite ions secreted by stimulated neutrophils in a few microliters of whole blood and have conducted clinical studies in humans. Here, we report normal reference intervals with our systems based on the results of 3,082 persons who underwent comprehensive cancer screening between February 2020 and March 2022. A total of 344 were extracted as reference individuals based on the results of the cancer screening and the reference intervals of the two systems were interim estimated considering gender and age. Reference intervals can be used as a marker of sub-clinical inflam­mation, which is difficult to detect with other blood markers.

Coenzyme Q10 is an essential lipid in the mitochondrial electron transport system and an important antioxidant. It declines with age and in various diseases, there is a need for a method to compensate for the decrease in coenzyme Q10. Resveratrol, a well-known anti-aging compound, has been shown to undergo metabolism to coenzyme Q10’s benzene ring moiety in cells. However, administration of resveratrol did not alter or only slightly increased total intracellular coenzyme Q10 levels in many cell types. Synthesis of coenzyme Q10 requires not only the benzene ring moiety but also the side chain moiety. Biosynthesis of the side chain portion of coenzyme Q10 is mediated by the mevalonic acid pathway. Here, we explore the impact of resveratrol on coenzyme Q10 levels in HepG2 cells, which possess a robust mevalonic acid pathway. As a results, intracellular coenzyme Q10 levels were increased by resveratrol admin­istration. Analysis using ¹³C₆-resveratrol revealed that the benzene ring portion of resveratrol was converted to coenzyme Q10. Inhibition of the mevalonic acid pathway prevented the increase in coenzyme Q10 levels induced by resveratrol administration. These results indicate that resveratrol may be beneficial as a coenzyme Q10-enhancing reagent in cells with a well-developed mevalonic acid pathway.

Drinking alcohol is considered one of the risk factors for develop­ment of diabetes mellitus. Recently, it was reported that selenoprotein P levels in blood are increased by ethanol intake. However, the mechanism by which ethanol increases selenoprotein P has not been elucidated. The expression of selenoprotein P protein and its mRNA were increased in a concentration- and time-dependent manner when human liver-derived HepG2 cells were treated with ethanol. Levels of AMPK and JNK proteins, which have been known to regulate selenoprotein P transcription, were unchanged by ethanol treatment. However, the amount of nuclear FoxO3a, a transcription factor of SeP, was increased. This was associated with dephosphorylation of ERK1 but not ERK2. It was found that ERK1 was dephosphorylated by activation of dual-specific phosphatase 5 and dual-specific phosphatase 6. However, the phosphorylation of MEK by ERK phosphokinase was not affected by ethanol treatment. These results suggest that the ethanol-induced increase in SeP levels occurs by enhanced tran­scrip­tion of SeP mRNA via the DUSP5/6–ERK1–FoxO3a pathway.

Types of fats and oils affect the onset of lifestyle diseases. In this study, we investigated the relationship between the postprandial hyperglycemia and fatty acids content in the skeletal muscle of C57BL/6 mice given 20% lard, palm oil, corn oil, safflower oil, and flaxseed oil for 16 weeks. Lard increased plasma glucose and insulin levels at the end of feeding period, whereas flaxseed oil did not. It was noteworthy that there is a positive correlation between palmitic acid content in the muscle and postprandial hyperglycemia, and a negative correlation between α-linolenic acid content and hyperglycemia. Alternatively, mice were given 30% lard for 16 weeks. When lard was partially substituted with flaxseed oil (10–50% substitution), flaxseed oil dose-dependently prevented lard-induced hyperglycemia and hyperinsulinemia. In conclusion, flaxseed oil prevents the adverse effects of lard through increasing in α-linolenic acid content in the muscle.

Neurological and skeletal muscle properties are suggested causes ‍of dynapenia. This study aimed to evaluate the relation­ship between upper limb muscle quality (grip strength/upper extremity muscle mass) and knee osteoarthritis in dynapenia, and to identify dynapenia-associated factors. Elderly individuals who responded to a public call for screening in Wakasa Town, Fukui Prefecture between June 2019 and November 2021 were included. The analysis included 433 participants (304 women aged 76.0 ‍± 7.1 years). Examination comprised (consecutively) a basic interview, physical function measurement, body composition measurement, and explanation of results. Dynapenia was observed in 67 patients. Binomial logistic regression analysis revealed that age, upper limb muscle quality score, and knee osteoarthritis were independent factors for dynapenia. Receiver operating charac­teristic analysis of the relationship between dynapenia and upper limb muscle quality showed an area under the curve of 0.806 (95% confidence interval: 0.658–0.953) for men (cut-off value, 14.3 ‍kg/kg) and 0.849 for women (95% confidence interval: 0.858–0.968; cut-off value, 14.0 ‍kg/kg). In conclusion, age, upper limb muscle quality, and knee osteoarthritis were independent factors of dynapenia. We demonstrated that upper limb muscle quality has good accuracy in detecting dynapenia in both men and women.

Nutritional information on hospitalized patients with COVID-19 is limited. We aimed to (1) investigate the prevalence of nutrition risk defined by the Scored Nutritional Risk Screening (NRS 2002) and malnutrition assessed by prognostic nutritional index (PNI) and controlling nutritional status score (CONUT), (2) observe the nutritional intervention, and (3) explore the predictors of critical condition and mortality. Nutritional risk was 53.00% and the prevalence of malnutrition was 79.09% and 88.79% among 464 patients based on PNI and CONUT, respectively. The area under the receiver operating characteristic curve for hypersensitivity C-‍reactive protein (hs-CRP), platelet-to-lymphocyte ratio (PLR), PNI, neutrophil/lymphocyte ratio (NLR), systemic immune-inflammation index (SII), and CONUT were 0.714, 0.677, 0.243, 0.778, 0.742, and 0.743, respectively, in discerning critical patients. The mortality-related area under the curve of hs-CRP, PLR, PNI, NLR, SII, and CONUT were 0.740, 0.647, 0.247, 0.814, 0.758, and 0.767, respectively. The results showed that CONUT and NLR were significantly correlated with the critical conditions. Our study revealed a high prevalence of nutritional risk and malnutrition among hospitalized patients with COVID-19. NLR, PLR, hs-CRP, SII, and CONUT are independent predictors of critical conditions and mortality. CONUT and NLR could assist clinicians in discerning critical cases.

In strategies to extend a healthy lifespan, early detection and prevention of frailty are critical. The purpose of this study was to analyze the current state and clinical risk factors of frailty among community-dwelling older to conduct a cross-sectional analysis of the individuals, correlation between frailty and nutrient intake, dietary diversity, and dietary patterns, and to elucidate the correlation between frailty-related dietary factors and the gut microbiota. The study included 786 participants aged ≥65 years from the Kyotango Multipurpose Cohort Study who had available data on their gut microbiota. Frailty was quantitatively assessed by selecting 32 items from the previously reported frailty index, with those scoring ≥0.21 classified as frailty (n = 119) and those with scores <0.21 as non-frailty (n = 667), followed by group comparisons. The frailty group had significantly higher values and rates than the non-frailty group for the following items: age, obesity (in females only), diabetes, hypertension, history of cancer treatment, polypharmacy, disturbed sleep quality, low physical activity, serum insulin levels, and high-sensitivity C-reactive protein. The frailty group had significantly lower levels of nutrients, including plant proteins, potassium (K), magnesium (Mg), iron (Fe), copper (Cu), vitamins B and C, folic acid, and total, soluble, and insoluble dietary fiber. When analyzed by food groups of dietary fiber, the frailty group had significantly lower intakes of soy products and non-green-yellow vegetables, specifi­cally. The Japanese Diet Index score (rJDI12) was significantly lower in the frailty group, with significant deficiencies in soy products and mushrooms included in the rJDI12. Cluster analysis of the Spearman correlation values between nutrient intake related to frailty and the gut microbiota abundance revealed a positive correlation between the cluster containing dietary fiber and the abundance of the phylum Bacillota, including the [Eubacterium]_eligens_group. In conclusion, our findings clarify the current state of frailty among older community residents and suggest the importance of a diverse range of plant-based foods, including soy products and non-green yellow vegetables, through correlation analysis with nutrients and food groups, and partially reveal the involvement of the gut microbiota.