Water-soluble and low molecular weight (Mw) β-glucans were successfully prepared by γ-irradiation of water-insoluble yeast β-glucans. The radiation dose used for the degradation of yeast β-glucan was remarkably reduced by increasing the pH of the sample or combining with hydrogen peroxide treatment. Radiation-degraded β-glucans with molecular weights in the range of 11−48 kDa reduced the total cholesterol, triglyceride, low density lipoprotein (LDL) cholesterol, and glucose levels in the blood of administered mice. The decreasing levels of both lipid and glucose indexes in the blood of tested mice strongly depended on the molecular weight of the β-glucan, and the radiation-degraded β-glucan with a molecular weight of about 25 kDa was found to be the most effective for the reduction of blood lipid and glucose levels. Particularly, the oral administration of 25 kDa β-glucan, with a daily dose of about 2 mg per head, reduced the total cholesterol, triglyceride, LDL-cholesterol, and glucose levels in the blood of tested mice to about 47.4%, 48.5%, 45.7%, and 47.2%, respectively. The effects on the reduction of blood lipid and glucose levels were also found to be stable after 20 days of stopping administration. These results indicate that the degraded β-glucan with a molecular weight of about 25 kDa prepared by γ-ray irradiation is a very promising ingredient that can be used in nutraceutical food for therapeutics of diabetic and dyslipidemia.

Vascular endothelial growth factor (VEGF) is involved in the growth of new blood vessels that feed tumors and kinesin spindle protein (KSP) plays a critical role in mitosis involving in cell proliferation. Simultaneous silencing of VEGF and KSP, an attractive and viable approach in cancer, leads on restricting cancer progression. The purpose of this study is to examine the therapeutic potential of dual gene targeted siRNA cocktail on human hepatocellular carcinoma Hep3B cells. The predesigned siRNAs could inhibit VEGF and KSP at mRNA level. siRNA cocktail showed a further downregulation on KSP mRNA and protein levels compared to KSP-siRNA or VEGF-siRNA, but not on VEGF expression. It also exhibited greater suppression on cell proliferation as well as cell migration or invasion capabilities and induction of apoptosis in Hep3B cells than single siRNA simultaneously. This could be explained by the significant downregulation of Cyclin D1, Bcl-2 and Survivin. However, no sigificant difference in the mRNA and protein levels of ANG2, involving inhibition of angiogenesis was found in HUVECs cultured with supernatant of Hep3B cells treated with siRNA cocktail, compared to that of VEGF-siRNA. Silencing of VEGF and KSP plays a key role in inhibiting cell proliferation, migration, invasion and inducing apoptosis of Hep3B cells. Simultaneous silencing of VEGF and KSP using siRNA cocktail yields promising results for eradicating hepatocellular carcinoma cells, a new direction for liver cancer treatment.

Background: Stem cell therapy for the treatment of vascular-related diseases through functional revascularization is one of the most important research areas in tissue engineering. The aim of this study was to investigate the in vitro differentiation of umbilical CL-MSC into endothelial lineage cells. Methods: In this study, isolated cells were characterized for expression of MSC-specific markers and osteogenic and adipogenic differentiation. They were induced to differentiate into endothelial-like cells and then examined for expression of the endothelial-specific markers, karyotype, and functional behavior of cells. Results: Isolated cells expressed MSC-specific markers and differentiated into adipocytes and osteoblasts. After endothelial differentiation, they expressed CD31, vWF, VE-cadherin, VEGFR1, and VEGFR2 at both mRNA and protein level, but their morphological changes were not apparent when compared with those of undifferentiated cells. There were no significant changes in karyotype of differentiated cells. Furthermore, angiogenesis assay and LDL uptake assay showed that differentiated cells were able to form the capillary-like structures and uptake LDL, respectively. Conclusion: The results indicated that umbilical CL-MSC could differentiate into functional endothelial-like cells. Also, they are suitable for basic and clinical studies to cure several vascular-related diseases.

The objective of this study was to evaluate replicative senescence of bovine granulosa cells (bGCs) during in vitro long-term culture. WST-1 assay analysis showed that bGCs proliferation was reduced from primary culture to 14th passage. The several bGCs from the 3rd passage and 7th passage exposed the weak activity of beta-galactosidase, while a strongly positive staining of beta-galactosidase was observed in bGCs from 14th passage. Flow cytometry analysis showed that bGCs were induced to cell cycle arrest at G0/G1 phase through in vitro expansion. TERT transcript expression of bGCs was downregulated from primary culture to 14th passage. The cell and nuclear area of bGCs were dramatically increased from 14th passage to 25th passage. The nucleocytoplasmic ratio of bGCs was dramatically reduced in 22th passage (4.32%) and 25th passage (2.45%), comparing to previous passages: primary culture (10.67%), 7th passage (9.21%), or 14th passage (10.33%). The number of microfilament bundle of bGCs was increased in 22nd passage (67.42 ± 17.76) and 25th passage (56.31 ± 22.45). The diameter of microfilament bundle of bGCs in 25th passage was dramatically increased to 1.88 ± 0.32 µm comparing to the primary culture (1.15 ± 0.03 µm). In this study, we also assessed the nuclear form factor which illustrates the level of nuclear circular form. A reduction of nuclear form factor was observed in bGCs during long-term in vitro expansion. The changes of nuclear form factor were correlated to other senescent characteristics, especially the nucleocytoplasmic ratio.

Simulated microgravity (SMG) induced the changes in cell proliferation and cytoskeleton organization, which plays an important factor in various cellular processes. The inhibition in cell cycle progression has been considered to be one of the main causes of proliferation inhibition in cells under SMG, but their mechanisms are still not fully understood. This study aimed to evaluate the effects of SMG on the proliferative ability and cytoskeleton changes of Chang Liver Cells (CCL-13). CCL-13 cells were induced SMG by 3D clinostat for 72 h, while the control group were treated in normal gravity at the same time. The results showed that SMG reduced CCL-13 cell proliferation by an increase in the number of CCL-13 cells in G0/G1 phase. This cell cycle phase arrest of CCL-13 cells was due to a downregulation of cell cycle-related proteins, such as cyclin A1 and A2, cyclin D1, and cyclin-dependent kinase 6 (Cdk6). SMG-exposed CCL-13 cells also exhibited a downregulation of α-tubulin 3 and β-actin which induced the cytoskeleton reorganization. These results suggested that the inhibited proliferation of SMG-exposed CCL-13 cells could be associate with the attenuation of major cell cycle regulators and main cytoskeletal proteins.