Functional Characterization and Structural Basis of an Efficient Di-C-glycosyltransferase from Glycyrrhiza glabra.

A highly efficient di-C-glycosyltransferase GgCGT was discovered from the medicinal plant Glycyrrhiza glabra. GgCGT catalyzes a two-step di-C-glycosylation of flopropione-containing substrates with conversion rates of >98%. To elucidate the catalytic mechanisms of GgCGT, we solved its crystal structures in complex with UDP-Glc, UDP-Gal, UDP/phloretin, and UDP/nothofagin, respectively. Structural analysis revealed that the sugar donor selectivity was controlled by the hydrogen-bond interactions of sugar hydroxyl groups with D390 and other key residues. The di-C-glycosylation capability of GgCGT was attributed to a spacious substrate-binding tunnel, and the G389K mutation could switch di- to mono-C-glycosylation. GgCGT is the first di-C-glycosyltransferase with a crystal structure, and the first C-glycosyltransferase with a complex structure containing a sugar acceptor. This work could benefit the development of efficient biocatalysts to synthesize C-glycosides with medicinal potential.

Baicalin Inhibits Coxsackievirus B3 Replication by Reducing Cellular Lipid Synthesis.

Baicalin is a flavonoid extracted from Scutellariae Radix and shows a variety of biological activities as reducing lipids, diminishing inflammation, and inhibiting bacterial infection. However, there is no report of baicalin against CVB3 infection. In this study, we found that baicalin can reduce viral titer in a dose-dependent manner in vitro at a dose with no direct virucidal effect. Moreover, we revealed that baicalin can also improve survival rate, reduce heart weight/body weight ratio, prevent virus replication, and relieve myocardial inflammation in the acute viral myocarditis mouse model induced by CVB3. Then, in order to explore the mechanism of baicalin inhibiting CVB3 replication, we respectively examined the expression of autophagosome marker LC3-II by Western blot, tested the concentration of free fatty acid (FFA) and cholesterol (CHO) by commercial kits, detected the mRNA levels of fatty acid synthase (Fasn) and acetyl coenzyme a carboxylase (ACC) by RT-PCR, and observed the lipid content of cells by fluorescence staining. The results showed that CVB3 infection increased autophagosome formation and lipid content in HeLa cells, but these changes were significantly blocked by baicalin. Finally, in order to confirm that baicalin inhibits viral replication and reduces autophagosome formation by reducing cellular lipids, we added exogenous palmitate to cell culture supernatants to promote intracellular lipid synthesis and found that palmitate did not alter LC3-II and CVB3/VP1 expression in HeLa cells with or without CVB3 infection. Interestingly, palmitate can reverse the inhibitory effect of baicalin on autophagosome formation and viral replication. In conclusion, our results indicated that lipids play an important role in CVB3 replication, and the effect of baicalin against CVB3 was associated with its ability to reduce cellular lipid synthesis to limit autophagosome formation.

The Effects of Photobiomodulation on MC3T3-E1 Cells via 630 nm and 810 nm Light-Emitting Diode.

BACKGROUND Photobiomodulation (PBM) has been explored as a promising therapeutic strategy to regulate bone cell growth; however, the effects of PBM on osteoblast cell lines remains poorly understood. In addition, as a light source of PBM, the light uniformity of light-emitting diode (LED) devices has not been given enough attention. MATERIAL AND METHODS Here, we sought to investigate the effects of PBM on MC3T3-E1 cells via 630 nm and 810 nm light from a newly designed LED with high uniformity of light. Cell proliferation, flow cytometric analysis, alkaline phosphatase (ALP) staining, ALP activity, Alizarin Red S staining, and quantitative real-time polymerase chain reaction (qRT-PCR) were carried out to assess treatment response. MC3T3-E1 cells were irradiated with LED devices (630±5 nm and 810±10 nm, continuous wave) for 200 seconds at a power density of 5 mW/cm² once daily. RESULTS Increases in cell proliferation and decreases in cell apoptosis were evident following irradiation. ALP staining intensity and activity were also significantly increased following irradiation. Level of mineralization was obviously enhanced in irradiated groups compared with non-irradiated controls. qRT-PCR also showed significant increases in mRNA expression of osteocalcin (OCN) and osteoprotegerin (OPG) in the irradiated groups. CONCLUSIONS Our results showed that LED PBM could promote the proliferation, ALP staining intensity and activity, level of mineralization, gene expression of OCN and OPG of MC3T3-E1 cells, with no significant difference between the 630 nm- and 810 nm-irradiated groups.

Emission characteristics of polycyclic aromatic hydrocarbons from combustion of different residential coals in North China.

Emission properties of polycyclic aromatic hydrocarbons (PAHs) from combustion of six residential coals in North China were investigated. The results indicated that, the total emission factors (EFs) for 15 PAH species in gaseous and particulate phases ranged from 52.8 to 1434.8 mg/kg with a decreasing sequence of local bituminous coals and anthracite coals, and honeycomb briquettes were largely dependent on the raw coals used to produce them. Particulate phase, dominated by median or high molecular weight components, made a major contribution (68.8%-76.5%) to the total EFs for bituminous coals, while gaseous phase with principal low molecular weight species accounted for most (86.3%-97.9%) of the total EFs for anthracite coals. The phase partitioning of PAH emission for honeycomb briquettes was similarly dependent on the crude coals. The total EFs, phase partitioning and component profiles of emitted PAHs were mainly influenced by the inner components of the studied coals. Burning mode and flue number on household coal-stoves also affected the emission characteristics by means of the oxygen supply. A sum of seven carcinogenic PAHs, benzo(a)pyrene(BaP)-equivalent carcinogenic power and total toxicity potency expressed in 2,3,7,8-tetrachlorodibenzo-dioxin(TCDD) toxic equivalence exhibited that bituminous coals and produced honeycomb briquettes had remarkably elevated values. Fluoranthene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene and indeno(1,2,3-cd)pyrene from anthracite coals showed higher levels of BaP-based toxic equivalent factor, though the other toxicity indices were rather low for this type of coal.

[Inhalation exposure assessment on gaseous PAHs in indoor air of folk houses in Beijing].

Seven gaseous polycyclic aromatic hydrocarbons (PAHs) were determined in the indoor air of 38 folk houses in Beijing during the heating and non-heating seasons, by the passive air samplers. The total average concentration of the gaseous PAHs was around 100 ng/m3, and the dominant components were those with 2 or 3 rings. The inhalation exposure of PAHs by the residents was calculated based on the concentrations of PAHs acquired, average house-staying time and respiratory rates at different ages. The calculated results indicated that, the potential total exposure rates of the seven gaseous PAHs for the adults (including senior people) during the heating and non-heating periods were 66 ng/h and 58 ng/h, respectively; while those for juvenile people were 56 ng/h and 50 ng/h, respectively.