Chemical composition of Galla chinensis extract and the effect of its main component(s) on the prevention of enamel demineralization in vitro

To determine the chemical composition of Galla chinensisextract （GCE） by several analysis techniques and to compare the efficacy of GCE and its main component（s） in inhibition of enamel demineralization, for the development of future anticaries agents, main organic composition of GCE was qualitatively determined by liquid chromatography-time of flight-mass spectrometry （LC-TOF-MS） and quantified by high-performance liquid chromatography-diode array detector （HPLC-DAD）. Inorganic ions were tested by inductively coupled plasma-atomic emission spectroscopy and F was especially measured by ion chromatography. Then, bovine enamel blocks were randomly divided into four treatment groups and were subjected to a pH-cycling regime for 12 times. Each cycle included 5-min applications with one of four treatments： 4g.L-1 GCE solution, 4g.L-1 gallic acid （GA） solution, I g.L-1 NaF solution （positive control）, deionized water （DDW, negative control）, and then 60-min application in pH 5.0 acidic buffer and 5-min application in neutral buffer. Acidic buffers were retained for calcium analysis. The main organic composition of GCE were GA and its isomer, and, to a lesser extent, small molecule gallotannins. The content of GA in GCE was 71.3%±0.2% （w/w）. Inorganic ions were present in various amounts, of which Ca was （136±2.82） μg.g-1, and Zn was （6.8±0.1） μg.g-1. No F was detected in GCE. in pH cycling, GA showed an effect similar to GCE in inhibiting enamel demineralization （P〉0.05）. GA was found to be the main effective, demineralization inhibiting component of GCE and could be a promising agent for the development of anticaries agents.

Effect of Galla chinensis on the remineralization of two bovine root lesions morphous in vitro

The present study aims to evaluate the effect of Galla chinensis compounds on the remineralization of two artificial root lesions morphous in vitro. Sixty bovine dentine blocks were divided into two groups and individually treated with two levels of demineralization solutions to form erosive and subsurface artificial carious lesions in vitro. Each group was then divided into three subgroups, each of which were treated with a remineralization solution （positive control）, deionized water （negative control）, or 4 000 mg-L-1 aqueous solutions of Galla chinensis extract. The dentine blocks were then subjected to a pH-cycling regime for 7 days. During the first 4 days, the daily cycle included 21-h deal and 3-h demineralization applications. The dentine blocks were dealt with the entire day during the remaining 3 days. Two specimens from each of the treatment groups were selected and observed under a polarized light microscope. Data collected using a laser scanning confocal microscope were computerized and analyzed. Galla chinensis extract clearly enhanced the remineralization of both erosive lesion and subsurface lesion patterns in the specimens （P〈0.05）. The level of remineralization of the erosive lesion by Galla chinensis extract was lower than that of the subsurface lesion （P〈0.05）. In addition, the remineralization of the subsurface lesion by Galla chinensis extract was higher than that of the remineralization solution （P〈0.05）. No significant difference between the remineralization of erosive lesions by Galla chinensis extract and the remineralization solution was observed （P〉0.05）. So Galla chinensis extract has the potential to improve the remineralization of artificial root lesions under dynamic pH-cyclic conditions, indicating its potential use as a natural remineralization medicine.

Effect of Galla chinensis on the In Vitro Remineralization of Advanced Enamel Lesions

Aim The effect of Galla chinensis on de-/re-mineralization of advanced enamel lesions was investigated by using micro-CT in a prolonged in vitro experiment. Methodology Baseline mineral contents of sound enamels were first analyzed. Then lesions were produced in an acidic buffer solution （2.2 mmol.L-1 Ca（NO3）2, 2.2 mmol-L1 KH2PO4, and pH=4.5） for 21 days, with thrice daily three-minute treatments, divided into four groups： Group A, 4 000 ppm crude aqueous extract of Galla chinensis （GCE）; Group B, 4 000 ppm gallic acid; Group C, 1 000 ppm F aq. （as NaF, positive control）; Group D, deionized water （negative control）. Next, the blocks were immersed in a remineralization solution （1.5 mmol.Lz CaC12, 0.9 mmol.L1 KH2PO4, 0.1 ppm F, and pH=7,0） for 200 days. Mineral loss （ML） in each region of interest （ROI） and integrated mineral loss （IML） of the lesions were calculated （comparing with baseline mineral content of sound enamel） at different time points. Results After 21 days demineralization, fluoride treatment showed a statistically significant demineralization-inhibiting effect among the four groups, and after 200 days of remineralization, mineral content recovery was ordered （lowest to highest） as A=C〈B〈D. Conclusion GCE could slow down the remineralization of enamel in the surface layer and thereby facilitate ion transport into the lesion body. The mechanism of Galla chinensis in enhancing the remineralization of dental caries is different from fluoride.

Determination of reactive oxygen generated from natural medicines and their antibacterial activity

Extracts of 16 natural medicine powders {Galla chinensis,Malloti cortex,Cassiae semen,Sophorae radix,Myricae cortex,Crataegi fructus,Gambit,Mume fructus,Geranii herba,Phellodendri cortex,Coptidis rhizoma,Swertiae herba,and Cinnamomi cortex） were assayed for reactive oxygen concentrations using the peroxyoxalate chemiluminescent detection system.High luminescence intensity was observed in Galla chinensis,Geranii herba,Malloti cortex,Myricae cortex,and Cinnamomi cortex.Additional experiments identified the reactive oxygen species as hydrogen peroxide.Galla chinensis generated 2.4 × 10^（-4） mol/L hydrogen peroxide from a 1 mg/mL solution.In bacterial growth tests,Galla chinensis extract had antibacterial activity against Escherichia coli.Staphylococcus aureus,Bacteroides thetaiotaomicron,Campylobacter sputorum biovar sputorum.Streptococcus salivarius thermophilus,Lactobacillus casei,and Bifidobacterium longum infantis.This antibacterial activity was decreased by the addition of catalase.It revealed that hydrogen peroxide which Galla chinensis produced participated in antibacterial activity.

基于比较转录组学的盐肤木响应五倍子蚜胁迫的机理探究

五倍子(Galla chinensis)是五倍子蚜寄生在盐肤木上形成的虫瘿,是中国常用的中药材之一。目前,关于五倍子形成过程中盐肤木对五倍子蚜的胁迫响应机制鲜有研究。本研究以成瘿及未成瘿的盐肤木叶片为材料,通过新一代转录组测序技术研究五倍子形成过程中盐肤木对五倍子蚜的胁迫响应机理。测序共获得成瘿组和未成瘿组间差异表达基因26 117个,其中上调基因21 581个,下调基因4 536个。通过GO和KEGG功能注释发现,差异基因主要富集在物质合成及代谢、遗传物质变化、植物激素及光合作用等途径。本研究共筛选到植物激素合成及信号传导途径差异表达基因292个。其中,乙烯(ET)合成及信号传导途径基因差异最显著,与胁迫响应密切相关,筛选到ET信号转导途径关键基因CTR1、MPK6在两种材料中表达量出现显著差异。此外,对筛选到的差异表达基因进行转录因子注释,共得到差异表达转录因子381个,其中83个与植物响应胁迫密切相关的转录因子家族基因表达量出现明显差异,包括MYB、bHLH、AP2-EREBP、NAC、WRKY、b ZIP。本研究通过二代高通量转录组测序技术研究五倍子蚜侵染下盐肤木胁迫响应相关基因的表达特征,为今后研究盐肤木响应五倍子蚜的胁迫机制及五倍子形成的分子机理奠定了基础。