莱鲍迪甙C高效转化细菌Paenarthrobacter ilicis CR5301全基因组测序及关键糖苷酶分析

为全面了解Paenarthrobacter ilicis CR5301的遗传背景,深入解析CR5301转化莱鲍迪甙C(rebaudioside C,RC)的关键酶,采用二代Illumina HiSeq和三代Nanopore相结合的测序方法,对CR5301进行全基因组测序和关键酶预测分析。结果显示,CR5301基因组为1个闭合环状染色体DNA分子,不含质粒,基因组序列全长4748281 bp,GC含量62.92%,共编码4458个基因,同时含有4个基因岛、1个前噬菌体和14个CRISPR-Cas编码序列。CR5301是P.ilicis物种第1个测定基因组完成图的菌株,也是Paenarthrobacter菌属已知基因组最大的菌株。基因组共线性分析和16S rRNA基因系统进化树分析结果一致表明,P.ilicis CR5301与P.aurescens具有更近的亲缘关系,而与P.ureafaciens的亲缘关系较远。7个蛋白质数据库综合注释分析发现,P.ilicis CR5301基因组共含有523个碳水化合物活性酶基因,其中18个糖苷酶基因可能是CR5301转化RC的关键酶基因。最后,通过ProtParam、SOPMA生物信息学软件,预测了18个糖苷酶的物化性质和二级结构。这些结果为CR5301的RC转化机制研究提供了清晰、完整的遗传信息,并且为P.ilicis物种广泛的生物学研究提供了完整、可靠的参考基因组序列,对今后P.ilicis物种生物学研究具有重要的参考价值和普遍的借鉴意义。

Radix Ilicis Pubescentis total flavonoids combined with mobilization of bone marrow stem cells to protect against cerebral ischemia/reperfusion injury

Previous studies have shown that Radix Ilicis Pubescentis total flavonoids have a neuroprotective effect, but it remains unclear whether Radix Ilicis Pubescentis total flavonoids have a synergistic effect with the recombinant human granulocyte colony stimulating factor-mobilized bone marrow stem cell transplantation on cerebral ischemia/reperfusion injury. Rat ischemia models were administered 0.3, 0.15 and 0.075 g/kg Radix Ilicis Pubescentis total flavonoids from 3 days before modeling to 2 days after injury. Results showed that Radix Ilicis Pubescentis total flavonoids could reduce pathological injury in rats with cerebral ischemia/reperfusion injury. The number of Nissl bodies increased, Bax protein expression decreased, Bcl-2 protein expression increased and the number of CD34-positive cells increased. Therefore, Radix Ilicis Pubescentis total flavonoids can improve the bone marrow stem cell mobilization effect, enhance the anti-apoptotic ability of nerve cells, and have a neuroprotective effect on cerebral ischemia/reperfusion injury in rats.

Radix Ilicis Pubescentis total flavonoids ameliorates neuronal damage and reduces lesion extent in a mouse model of transient ischemic attack

Flavonoids are a major component in the traditional Chinese medicine Radix Ilicis Pubescentis.Previous studies have shown that the administration of Radix Ilicis Pubescentis total flavonoids is protective in cerebral ischemia.However,to our knowledge,no studies have examined whether the total flavonoids extracted from Radix Ilicis Pubescentis prevent or ameliorate neuronal damage following transient ischemic attacks.Therefore,Radix Ilicis Pubescentis total flavonoids question and the potential underlying mechanisms.Thus,beginning 3 days before the induction of a mouse model of transient ischemic attack using tert-butyl hydroperoxide injections,mice were intragastrically administered 0.3,0.15,or 0.075 g/kg of Radix Ilicis Pubescentis total flavonoids daily for 10 days.The results of spectrophotometric analyses demonstrated that Radix Ilicis Pubescentis total flavonoids enhanced oxygen free radical scavenging and reduced pathological alterations in the brain.Hematoxylin-eosin staining results showed that Radix Ilicis Pubescentis total flavonoids reduced hippocampal neuronal damage and cerebral vascular injury in this mouse model of transient ischemic attack.These results suggest that the antioxidant effects of Radix Ilicis Pubescentis total flavonoids alleviate the damage to brain tissue caused by transient ischemic attack.

Effect of Radix Ilicis Pubescentis on the Rats with Hyperlipidemia

[Objectives]To study the hypolipidemic effect of Radix Ilicis Pubescentis and to provide experimental scientific basis for the study of hypolipidemic mechanism of Radix Ilicis Pubescentis.[Methods]60 healthy(SD)male rats were selected,10 rats were given basic feed and pure water as blank control group,and the other 50 rats were fed with high-fat diet for 4 weeks.After establishing the hyperlipidemic rat model,the rats were randomly divided into model group,high,middle and low dose groups(1000,800,500 mg/kg),and positive control group(simvastatin 0.4 mg/mL).The corresponding drugs were intragastrically administered to the rats in each group(blank control group and model group intragastrically administered with normal saline).Four weeks later,blood samples from abdominal aorta were taken to detect serum total cholesterol(TC),serum triglyceride(TG),low density lipoprotein cholesterol(LDL-C),and high density lipoprotein cholesterol(HDL-C).[Results]Compared with the blank control group,TC,TG and LDL-C increased while HDL-C decreased in the model group(P<0.05);compared with the model group,TC,TG and LDL-C in Radix Ilicis Pubescentis group decreased to varying degrees,and the content of HDL-C in serum of rats had no statistical significance.Compared with the blank control group,the serum TC and TG contents of the model group,positive control group,high,middle and low dose groups of Radix Ilicis Pubescentis extract significantly increased(P<0.01 or 0.05),and the serum LDL-C content of rats decreased(P<0.05).[Conclusions]The extract of Radix Ilicis Pubescentis had a certain lipid-lowering effect on hyperlipidemia in rats,and had obvious preventive and therapeutic effect on hyperlipidemia.

Research Overview of Zhuang Medicine Fumigation Lotions

At present,there is little research on the application of Zhuang medicine fumigation lotions in skin diseases,and there is no research on related preparation processes and quality standards.In this paper,the characteristics of its formulation and the research progress in skin diseases are reviewed,in order to deepen the understanding of Zhuang medicine fumigation lotions and provide a more detailed theoretical basis for its clinical research,development and application.

Quantitative determination of ilexgenin A in rat plasma by liquid chromatography coupled with mass spectrometry and its pharmacokinetics

A sensitive and selective high performance liquid chromatography coupled with electrospray ionization mass spectrometry （LC-MS） was developed for the quantitative determination of ilexgenin A （IA）,a major component in Radix Ilicis Pubescentis,in rat plasma.Chromatographic separation was performed on a C 18 column,with methanol-5 mM ammonium acetate （80：20,v/v） as the mobile phase.Mass spectrometer was set in negative mode with target ions at m/z 501.1→501.1 for IA and m/z 779.4→779.4 for digoxin （internal standard,IS）.Rat plasma was extracted with ethyl acetate after addition of phosphoric solution and the organic layer was evaporated and reconstituted with mobile phase for LC-MS analysis.The proposed method was validated with a linear range of 1.05-525.5 ng/mL for IA with limit of quantitation （LOQ） at 1.05 ng/mL.Intra-and inter-day precision expressed as relative standard deviation （RSD） were less than 10% at LOQ level and overall recovery was over 80%.This validated method was used successfully for the pharmacokinetic study of IA in rats after oral dosing of IA （100 mg/kg） and some main pharmacokinetic parameters of IA in rats were obtained.