Soybean processing, e.g., by soaking, heating, and fermentation, typically results in diverse metabolic changes. Herein, multivariate analysis-based metabolic profiling was employed to investigate the effects of fermentation by Aspergillus oryzae or Bacillus subtilis on soybean substrates extracted at 4, 25, or 55 °C. As metabolic changes for both A. oryzae and B. subtilis were most pronounced for substrates extracted at 55 °C, this temperature was selected to compare the two microbial fermentation strategies, which were shown to be markedly different. Specifically, fermentation by A. oryzae increased the levels of most organic acids, γ-aminobutyric acid, and glutamine, which were ascribed to carbohydrate metabolism and conversion of glutamic acid into GABA and glutamine. In contrast, fermentation by B. subtilis increased the levels of most amino acids and isoflavones, which indicated the high activity of proteases and β-glucosidase. Overall, the obtained results were concluded to be useful for the optimization of processing steps in terms of nutritional preferences.

The roots of Salvia miltiorrhiza are known to exhibit antioxidant and antibacterial activities. To improve the antioxidant and antibacterial activities of S. miltiorrhiza roots, the roots were fermented with Aspergillus oryzae at 25 °C for 3 weeks. The non-fermented (SME) and fermented (SMBE) roots of S. miltiorrhiza were extracted with 70% ethanol, respectively, and then fractionated with organic solvents. By fermentation, total phenolic and flavonoid contents, as well as antioxidant activity of SMBE, were increased by about 1.2 to 1.3 times compared with those of SME. The antibacterial activity of SMBE was also twice as high as that of SME. The antibacterial activity of SMBE against Bacillus cereus was lower in the n-hexane and chloroform fractions, but higher in the ethyl acetate and n-butanol fractions, compared with those of SME. These results indicate that the bioactive components of S. miltiorrhiza roots exhibiting antibacterial activity were converted to more polar compounds by fermentation of A. oryzae. Gas chromatography and mass spectrometry (GC-MS) and LC-MS analyses of SME and SMBE demonstrate that these changes are due to the acylation of dihydrofuran-2(3H)-one, dealkylation of 4-methylbenzene-1,2-diol and 4-ethylbenzene-1,2-diol, and esterification of hexadecanoic acid and (9Z, 12Z)-octadec-9,12-dienoic acid during fermentation.

In this study, bisphasic calcium phosphate (BCP) and two types of polysaccharide, carboxymethyl cellulose (CMC) and hyaluronic acid (HyA), were used to fabricate composite block bone grafts, and their physical and biological features and performances were compared and evaluated in vitro and in vivo. Specimens of the following were prepared as 6 mm diameter, 2 mm thick discs; BPC mixed with CMC (the BCP/CMC group), BCP mixed with crosslinked CMC (the BCP/c-CMC group) and BCP mixed with HyA (the BCP/HyA group) and a control group (specimens were prepared using particle type BCP). A scanning electron microscope study, a compressive strength analysis, and a cytotoxicity assessment were conducted. Graft materials were implanted in each of four circular defects of 6 mm diameter in calvarial bone in seven rabbits. Animals were sacrificed after four weeks for micro-CT and histomorphometric analyses, and the findings obtained were used to calculate new bone volumes (mm³) and area percentages (%). It was found that these two values were significantly higher in the BCP/c-CMC group than in the other three groups (p < 0.05). Within the limitations of this study, BCP composite block bone graft material incorporating crosslinked CMC has potential utility when bone augmentation is needed.

Malva verticillata, also known as Chinese mallow, is an herbaceous plant with colorful flowers and has been used as a medicine for thousands of years. This study investigated this herb for potential antioxidant activity or an association with osteoblast differentiation. M. verticillate leaves were fermented with B. subtilis MV1 at 30 °C for 7 days to enhance their biological activities. The resultant aqueous extract (MVW) and the fermented leaves (MVB) were measured for antioxidant and osteoblast differentiation. The results showed that the total phenolic, flavonoid, and antioxidant activity, as well as the osteoblast differentiation of the MVB increased (2 to 6 times) compared with those of the MVW. MVB induced phosphorylation of p38, extracellular signal-regulated kinase in C3H10T1/2 cells, and the phosphorylation was attenuated via transforming growth factor-β (TGF-β) inhibitors. Moreover, runt-related transcription factor 2 and osterix in the nucleus increased in a time-dependent manner. The messenger RNA expression of alkaline phosphatase and bone sialoprotein increased about 9.4- and 65-fold, respectively, compared to the non-treated cells. MVB stimulated C3H10T1/2 cells in the osteoblasts via TGF-β signaling. Thus, fermented M. verticillata extract exhibited enhanced antioxidant activity and osteoblast differentiation.

In this study, hydroxypropyl methylcellulose (HPMC) was mixed with particle-type xenografts, derived from two different species (bovine and porcine), to increase the manipulability of bone grafts and compare the bone regeneration ability. Four circular defects with a diameter of 6 mm were formed on each rabbit calvaria, and the defects were randomly divided into three groups: no treatment (control group), HPMC-mixed bovine xenograft (Bo-Hy group), and HPMC-mixed porcine xenograft (Po-Hy group). At eight weeks, micro-computed tomography (µCT) scanning and histomorphometric analyses were performed to evaluate new bone formation within the defects. The results revealed that the defects treated with the Bo-Hy and the Po-Hy showed higher bone regeneration than the control group (p < 0.05), while there was no significant difference between the two xenograft groups (p > 0.05). Within the limitations of the present study, there was no difference in new bone formation between porcine and bovine xenografts with HPMC, and bone graft material was easily moldable with the desired shape during surgery. Therefore, the moldable porcine-derived xenograft with HPMC used in this study could be a promising substitute for the currently used bone grafts as it exhibits good bone regeneration ability for bony defects.

Aralia continentalis has been used in Korea as a folk remedy for arthralgia, rheumatism, and inflammation. However, its anti-lymphoma effect remains uncharacterized. Here, we demonstrate that A. continentalis extract and its three diterpenes efficiently kill B-lymphoma cells. Our in vitro and in vivo results suggest that the cytotoxic activities of continentalic acid, a major diterpene from A. continentalis extract, are specific towards cancer cells while leaving normal murine cells and tissues unharmed. Mechanistically, continentalic acid represses the expression of pro-survival Bcl-2 family members, such as Mcl-1 and Bcl-xL. It dissociates the mitochondrial membrane potential, leading to the stimulation of effector caspase 3/7 activities and, ultimately, cell death. Intriguingly, this agent therapeutically synergizes with roflumilast, a pan-PDE4 inhibitor that has been successfully repurposed for the treatment of aggressive B-cell malignancies in recent clinical tests. Our findings unveiled that A. continentalis extract and three of the plant’s diterpenes exhibit anti-cancer activities. We also demonstrate the synergistic inhibitory effect of continentalic acid on the survival of B-lymphoma cells when combined with roflumilast. Taken in conjunction, continentalic acid may hold significant potential for the treatment of B-cell lymphoma.