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Agarase and its metabolites are reported to have applications in a variety of fields, but there have been few studies of the effects of agaro-oligosaccharide hydrolysate on muscle function. In this study, we analyzed the functionality of agarase and its metabolites in bacteria isolated from seawater. A bacterium with agar-degrading activity was isolated from Shimabara, Nagasaki, Japan. Through 16S rRNA sequence alignment, it was identified as being closely related to Rheinheimera sp. WMF-1 and was provisionally named Rheinheimera sp. (HY). Crude enzymes derived from this bacterium demonstrated an ability to hydrolyze various polysaccharides, including agar, agarose, and starch, with the highest specificity observed for agarose. The optimum pH and temperature were pH 10 and 50 °C. A glycoside bond specificity analysis of enzymatic activity indicated the cleavage of the α-linkage. Next, we investigated the functional effects of agaro-oligosaccharides on C2C12 myotubes. Treatment with 10-30 kDa oligosaccharides significantly increased the hypertrophy rate, diameter, and expression of myosin heavy-chain genes in C2C12 myotubes. These results indicate that the agaro-oligosaccharides produced by the enzymes identified in this study improve muscle mass, suggesting their potential contribution to muscle function.

Chaga mushroom, Inonotus obliquus, was used as food and nutrient food and traditional herbs in Russia, China and Japan, with anti-inflammatory and anticancer activities. Chemical investigations of the fruiting bodies of Chaga were carried to uncover the bioactive metabolites. As a result, seven undescribed lanostane-type triterpenoids, namely inonotusols H-N, were isolated, and all lanostanoids remarkably inhibited NO production in lipopolysaccharide-stimulated BV-2 microglial cells. Of these, inonotusols I and L presented the most potent inhibitory effects on inducible nitric oxide synthase (iNOS) and NO production without any significant cytotoxicity. Molecular docking studies confirmed the capacity of inonotusols I and L to interact with iNOS protein. Structure-activity relationships were also discussed. These results indicated that the potential anti-inflammatory effects of inonotusols I and L in microglial BV-2 cells may be imparted through suppression of iNOS. These results may support the use of I. obliquus for food and medicinal application., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Ethnopharmacological Relevance: Calculus bovis (C. bovis), a widespread known traditional animal drug in China and Japan, has been widely used for a long time to treat various diseases, including high fever, convulsion and stroke. The aim of the present paper is to comprehensively review knowledge about C. bovis in terms of traditional usages, origin, chemical constituents, pharmacological activities and toxicology to seek an applicable substitute for NCB and provide potential new strategies utilizing C. bovis. Additionally, directions and perspectives for future investigations regarding C. bovis are also discussed., Materials and Methods: In this paper, the traditional usages, origin, chemical constituents, pharmacology, and toxicology of C. bovis are comprehensively and systematically summarized by searching scientific databases, including Web of Science, PubMed, ScienceDirect, Springer, CNKI, Baidu Scholar and others. Additionally, some classic books of Chinese herbal medicine, academic papers authored by individuals with MSc and PhD degrees, local government reports as well as the state of local drug standards are also retrieved., Results: Currently, C. bovis mainly derives from four sources: natural Calculus bovis (NCB), Calculus bovis sativus (CBS), Cultured calculus bovis (CCB) and Calculus bovis artifactus (CBA). Owing to their different formation processes, the chemical constituents of the four kinds of C. bovis show certain differences. Additionally, over 44 chemical constituents have been isolated and identified from C. bovis, mainly including bile pigments, bile acids, cholesterols and amino acids. Further investigations have revealed a wide range of pharmacological effects of C. bovis, with effects on the nervous system, cardiovascular system, respiratory system, digestive system, immune system and others. Furthermore, NCB and CBA show hypotoxicity, but high concentrations of bilirubin can cause neurotoxicity and hearing impairment. Additionally, pharmacokinetic data for C. bovis are still lacking., Conclusion: CBS contains analogous types and amounts of constituents and exerts similar therapeutic effects to NCB. Thus, CBS might be used as a sustainable substitute for NCB. Furthermore, the configuration and concentration of bile acids and bilirubin in C. bovis are responsible for the difference in pharmacological effects in the four types C. bovis. Further studies should focus on the structure-function relationship of bile acids and bilirubin in C. bovis by employing pharmacokinetics., Competing Interests: Declaration of competing interest The authors declare there are no conflicts of interest. This manuscript has not been published or accepted elsewhere. It is not being submitted to any other journal. We declare that the present study was performed according to international, national and institutional rules considering animal experiments, clinical studies and biodiversity rights., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: The role of adjuvant radiotherapy for positive lymph nodes (LN) in patients with esophageal cancer who received neoadjuvant concurrent chemoradiotherapy (CCRT) is not well established. This study is aimed at determining the impact of positive LN and the survival benefit of postoperative radiotherapy (PORT) after CCRT plus surgery on esophageal cancer patients., Methods: Seventy patients with positive LN after neoadjuvant CCRT followed by esophagectomy were enrolled in the study. Patients were grouped into surgery alone following neoadjuvant CCRT (n = 41) and surgery plus PORT following neoadjuvant CCRT (n = 29) groups. The preoperative radiation dose was 36-45 Gy (mean 40 Gy) and the postoperative radiation dose was 20 Gy in 10 fractions., Results: The 5-year survival rate and mean survival was 40% and 58.6 ± 53.9 months for patients with a pathologic complete response (pCR) compared with 8.3% and 22.7 ± 35.5 months, respectively, for non-pCR patients (p = 0.026). Local and distant recurrent patterns were similar for patients who did and did not receive PORT (p = 0.876). The mean survival did not differ significantly between the 2 groups (p = 0.889). Pathological complete response to CCRT was the only significant factor influencing survival (p = 0.026)., Conclusions: Postoperative RT did not improve survival in patients with positive LN after CCRT followed by curative surgery for esophageal cancer., (© 2017 S. Karger AG, Basel.)

Balanophora japonica and B. yakushimensis are two putatively agamospermic taxa previously reported from southern Japan. Their inflorescences superficially represent those of B. laxiflora and B. fungosa. In this study we confirmed their presence in Taiwan by morphological and phylogenetic analysis using nuclear 18S rDNA and nrITS sequences with related taxa. B. japonica, B. yakushimensis, and B. laxiflora formed a well-supported clade that is distinct from other Balanophora. All three taxa also show considerable differences on morphological and nucleotide sequence differences, therefore the name of B. yakushimensis is retained. The results provide new insights on the intrageneric classification of Balanophora and suggest the positioning of female flowers should be down-weighted. We also successfully identify the hosts of B. japonica and B. yakushimensis by amplifying chloroplast matK sequences from the connected root tissues. The results showed that B. japonica parasitizes on Symplocos species, and that B. yakushimensis parasitizes on Distylium racemosum in Japan and Schima superba in Taiwan's population.