AIM:To investigate the effect of different low-dose mifepristone on apoptosis in granulosa cells and to test low-dose mifepristone as an orally contraceptive drug.METHODS:By using immunofluorescence,terminal deoxynucleotidyl transferase-mediated nick end labelling(TUNEL)and flow cytometry technique,the nuclear morphologic features and ratio of apoptosis and fluorescent intensity of caspase-3 in granulosa cells cultured in vitro treated with different low-doses of mifepristone were observed,respectively.RESULTS:By the display of immunofluorescence,the granulosa cells in treatment group were classified as apoptotic cells on the basis of their morphologic features contained a single condensed chromatin,multiple nuclear fragments.The results of TUNEL showed significant difference between control group and groups treated with different concentration of mifepristone(P

Objective To explore the effects of low-dose mifepristone on human granulosa cells and the molecular mechanism of its inhibiting or delaying ovulation.Methods Phase contrast microscopy and electron microscopy were conducted to observe the morphological and biochemical change of granulosa cells exposed to various doses of mifepristone for 24 hours(the final concentrations of mifepristone were respectively 2.5?mol/L,5?mol/L and 10?mol/L,Non-mifepristone as the control group).Immunocytochemistry was performed to detect the protein expressions of Bcl-2 and Bax in granulosa cells exposed to various doses of mifepristone for 24 hours.The mRNA levels of Bcl-2 and Bax in granulosa cells were detected by RT-PCR.Results Granulosa cells treated with various doses of mifepristone presented typical morphological characteristics of apoptosis such as condensation of chromatin and cytosolic,and shrink of cells.The expression of Bcl-2 protein might be down-regulated,while the expression of Bax might be up-regulated by mifepristone.The histological score(HSCORE) of Bcl-2 protein in granulosa cells,which were incubated with various doses of mifepristone(2.5?mol/L,5?mol/L,10?mol/L) for 24 hours,were respectively 2.15?0.16,1.88?0.13 and 1.64?0.16 respectively,Compared with the HSCORE of the control group(2.51?0.16),the difference was significant(P0.05).Conclusion Mifepristone could induce human granulosa cells apoptosis.The Bcl-2 protein and Bax protein may be involved in the apoptosis of granulosa cells induced by mifepristone.

Clustered regularly interspaced short palindromic repeats (CRISPR) is an acquired immune system existing in archaea and bacteria with the long-term process of evolutionary.CRISPR/Cas9 gene editing system is a new type of gene editing technology developed based on the system.CRISPR/Cas9 is a more efficient method for gene targeting than the previous methods.It has been successfully applied for gene-modified of eukaryotes since 2012,but the reports about pathogenic microorgaisms are rarely.Here,the research progress in the structure,mechanism of CRISPR/Cas9 system and its applications on pathogenic microorgaisms is reviewed.

Decoction is one of the most commonly used dosage forms of traditional Chinese medicine. The stability of chemical constituents in decoction is closely related to the clinical efficacy and safety. There were few reports about the influence of metal ions in the stability of chemical constituents in traditional Chinese medicine. However, there is no evidence that metal ions in decoction water need to be controlled. In this study, 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (THSG), one of the main constituents in Polygoni Multiflori Radix was studied. Ordinary tap water, deionized water, and water containing different metal ions were used to investigate and compare the influence on THSG. The results showed that after storage in a dark place at the room temperature for 10 days, the degradation of THSG was 7% in deionized water, while undetectable in tap water. The content of THSG could be decreased by different kinds of metal ions, and the effect was concentration-dependent. Moreover, Fe3+ and Fe2+ showed the greatest influence at the same concentration; and our study has shown that THSG decreased more than 98% in Fe and Fe2+ solutions at 500 ppm concentration. In the same time we found out p-hydroxybenzaldehyde (molecular weight: 122.036 7) and 2,3,5-trihydroxybenzaldehyde-2-O-glycoside (molecular weight: 316.079 4) were the main degradation products of THSG in tap water and water containing Cu2+, Ca2+, Zn2+, Mg2+ and Al3+. The product of THSG dimer with a water molecule was found in water containing Fe3+ and Fe2+. The above results showed that the metal ions in water could significantly influence the stability of THSG in water, indicating that the clinical efficacy and safety of decoction would be affected if the metal ions in water were not under control. It's suggested that deionized water should be used in the preparation of decoction containing Polygoni Multiflori Radix in the clinic to avoid degradation of THSG. Meanwhile, decoction prepared by tap water should be taken by patients in a short time. Our investigation provides important information and reference about the influence of metal ions on the stability of decoctions in other traditional Chinese medicine that have unstable groups such as hydroxyls and unsaturated bonds, etc.