Objective To evaluate the effect of different honey processing methods on the dissolution of Cynanchum salicifolia in vitro.Methods High performance liquid chromatography ① Carotene and β-Determination method of sitosterol content:chromatographic column:Dia monosil C18(250 mm×4.6 mm,5 μm).Mobile phase:methanol:0.05％phosphoric acid water=99∶1.Isocratic elution;Flow rate 1.0 mL·min-1;Detection wavelength 209 nm;Injection volume 10 μL.The column temperature is 30℃.②Determination method of glucose and fructose:the chromatographic column is Inspire NH2 sugar analysiscolumn(250 mm×4.6 mm,5 μm).The mobile phase was acetonitrile water(77∶23).The flow rate is 0.8 mL·min-1;The column temperature is 35℃;Injection volume 10 μL;Evaporative light scattering detector(ELSD):the drift tube temperature is 100℃,and the carrier gas flow rate is 2.8 L·min-1.Results ①The results showed that carotene and β-the determination of sitosterol showed a good linear relationship in the range of mass concentration(r＞0.9999).The average recoveries were 99.71％and 98.83％.The RSD of precision,stability and repeatability were all less than 2.0％.②The established method of glucose fructose showed a good linear relationship in the range of 0.05-0.50 mg·mL-1(r=0.9991).Conclusion The content determination method and in vitro dissolution test methodology of the study meet the requirements,which provides a scientific basis for the establishment of quality standards and modern research of honey roasted Paeonia salicifolia.

Objective To evaluate the effect of different honey processing methods on the dissolution of Cynanchum salicifolia in vitro.Methods High performance liquid chromatography ① Carotene and β-Determination method of sitosterol content:chromatographic column:Dia monosil C18(250 mm×4.6 mm,5 μm).Mobile phase:methanol:0.05％phosphoric acid water=99∶1.Isocratic elution;Flow rate 1.0 mL·min-1;Detection wavelength 209 nm;Injection volume 10 μL.The column temperature is 30℃.②Determination method of glucose and fructose:the chromatographic column is Inspire NH2 sugar analysiscolumn(250 mm×4.6 mm,5 μm).The mobile phase was acetonitrile water(77∶23).The flow rate is 0.8 mL·min-1;The column temperature is 35℃;Injection volume 10 μL;Evaporative light scattering detector(ELSD):the drift tube temperature is 100℃,and the carrier gas flow rate is 2.8 L·min-1.Results ①The results showed that carotene and β-the determination of sitosterol showed a good linear relationship in the range of mass concentration(r＞0.9999).The average recoveries were 99.71％and 98.83％.The RSD of precision,stability and repeatability were all less than 2.0％.②The established method of glucose fructose showed a good linear relationship in the range of 0.05-0.50 mg·mL-1(r=0.9991).Conclusion The content determination method and in vitro dissolution test methodology of the study meet the requirements,which provides a scientific basis for the establishment of quality standards and modern research of honey roasted Paeonia salicifolia.

Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Humans ; Apolipoprotein E4/genetics* ; Cytosine ; Mutation ; Blastocyst ; Heterozygote ; Gene Editing ; CRISPR-Cas Systems