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.

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

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.

Objective:To investigate the risk factors for lower extremity deep vein thrombosis (DVT) in patients with bone trauma and their diagnostic efficacy.Methods:A retrospective cohort study was conducted to analyze the clinical data of 108 patients with bone trauma who were admitted to Affiliated Changzhou Second People′s Hospital of Nanjing Medical University from October 2023 to February 2024, including 61 males and 47 females, aged 17-96 years [(55.2±19.5)years]. Based on the results of color Doppler ultrasonography of lower extremities within 96 hours on admission, the patients were divided into DVT group ( n=58) and non-DVT group ( n=50). In DVT group, 42 patients developed lower extremity DVT within 7 days after trauma and the other 16 patients developed lower extremity DVT after 7 days. Basic clinical data including gender, age, body mass index (BMI), underlying diseases, cause of injury, site of fracture, surgery and admission Caprini score, and admission laboratory test indicators including routine coagulation indicators [prothrombin time (PT), international normalized ratio (INR), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (FBG) and D-dimer (D-D)] and four thrombosis indicators [plasma thrombin-antithrombin III complex (TAT), thrombomodulin (TM), tissue-type plasminogen activator-inhibitor 1 complex (tPAIC) and plasmin-alpha2-plasmin inhibitor complex (PIC)] were collected in the two groups. Univariate analysis and multivariate binary Logistic regression analysis were conducted to investigate the correlation between these indicators and incidence of lower extremity DVT in patients with bone trauma and determine the independent risk factors. Receiver operating characteristic (ROC) curve and area under the curve (AUC) of the relevant risk factors were analyzed to evaluate and compare the diagnostic efficacy of the factors for lower extremity DVT in patients with bone trauma and further assess the diagnostic efficacy of the factors for lower extremity DVT within 7 days after bone trauma. Results:Univariate analysis revealed significant correlations of gender, age, Caprini score, D-D, TAT, TM and PIC with incidence of lower extremity DVT in patients with bone trauma ( P<0.01). The results of multivariate binary Logistic regression analysis demonstrated that Caprini score ( OR=1.36, 95% CI 1.12, 1.65, P<0.01), TAT ( OR=1.05, 95% CI 1.00, 1.10, P<0.05), and TM ( OR=1.34, 95% CI 1.02, 1.77, P<0.05) were significantly correlated to incidence of lower extremity DVT in patients with bone trauma. ROC curve analysis indicated that TAT (AUC=0.76, 95% CI 0.67, 0.86) had the highest diagnostic efficiency, followed by TM (AUC=0.72, 95% CI 0.62, 0.81) and Caprini score (AUC=0.72, 95% CI 0.62, 0.82). The combined analysis of all the factors effectively enhanced the diagnostic efficiency for DVT (AUC=0.84, 95% CI 0.77, 0.92). Additionally, TAT (AUC=0.81, 95% CI 0.71, 0.91) demonstrated better diagnostic efficacy for lower extremity DVT within 7 days after bone trauma compared with the Caprini score (AUC=0.72, 95% CI 0.61, 0.83) and TM (AUC=0.71, 95% CI 0.60, 0.83). Similarly, the combined analysis of all the factors also effectively enhanced the overall diagnostic efficacy for lower extremity DVT within 7 days after bone trauma (AUC=0.85, 95% CI 0.77, 0.93). Conclusions:Caprini score, TAT and TM are identified as independent risk factors for lower extremity DVT in patients with bone trauma, and all the three factors demonstrate good diagnostic efficacy. Their combination is found to have statistically significant higher diagnostic efficiency than each individual factor. Furthermore, TAT is proved to be the best in diagnosing lower extremity DVT within 7 days after bone trauma, while the combined analysis of all the risk factors can further improve the diagnostic efficacy.

The main constituents of a typical medicinal herb,Polygonum muMflorum (Heshouwu in Chinese),that induces idiosyncratic liver injury remain unclear.Our previous work has shown that cotreatment with a nontoxic dose of lipopolysaccharide (LPS) and therapeutic dose of Heshouwu can induce liver injury in rats,whereas the solo treatment cannot induce observable injury.In the present work,using the constituent "knock-out" and "knock-in" strategy,we found that the ethyl acetate (EA) extract of Heshouwu displayed comparable idiosyncratic hepatotoxicity to the whole extract in LPS-treated rats.Results indicated a significant elevation of plasma alanine aminotransferase,aspartate aminotransferase,and liver histologic changes,whereas other separated fractions failed to induce liver injury.The mixture of EA extract with other separated fractions induced comparable idiosyncratic hepatotoxicity to the whole extract in LPS-treated rats.Chemical analysis further revealed that 2,3,5,4'-tetrahydroxy trans-stilbene-2-O-β-glucoside (trans-SG) and its cis-isomer were the two major compounds in EA extract.Furthermore,the isolated cis-,and not its trans-isomer,displayed comparable idiosyncratic hepatotoxicity to EA extract in LPS-treated rats.Higher contents of cis-SG were detected in Heshouwu liquor or preparations from actual liver intoxication patients associated with Heshouwu compared with general collected samples.In addition,plasma metabolomics analysis showed that cis-SG-disturbing enriched pathways remarkably differed from trans-SG ones in LPS-treated rats.All these results suggested that cis-SG was closely associated with the idiosyncratic hepatotoxicity of Heshouwu.Considering that the cis-trans isomerization of transSG was mediated by ultraviolet light or sunlight,our findings serve as reference for controlling photoisomerization in drug discovery and for the clinical use of Heshouwu and stilbene-related medications.