We developed a detailed electroporation method to deliver efficiently siRNA into mouse preimplantation embryos. By introducing Cy3 labeled negative control small interfering RNA (siRNA) into mouse preimplantation embryos, we optimized conditions for the electroporation, including the voltage, pulse duration, pulse number, electroporation buffer and an important step to weaken the zona pellucida. Embryonic survival rate, transfection rate and blastocyst development rate were evaluated under the converted fluorescence microscope, by embryos counting and statistical analysis. The best transfection was achieved in opti-MEM under the conditions of 30 V, 1 ms, 3 pulses, and the duration of digestion in tyrode's solution was 10 s. In conclusion, the proposed electroporation approach here is a simple and efficient tool to deliver siRNA for RNA interference (RNAi) into mouse preimplantation embryos.
Animals ; Blastocyst ; metabolism ; Electroporation ; Female ; Male ; Mice ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; methods

OBJECTIVE To investigate the reasons for interrupting menopausal hormone therapy in patients with menopausal syndrome and analyze the influencing factors that may lead to treatment interruption. METHODS The patients who visited our menopause medicine clinic from March 2022 to November 2023 and established a menopausal health manual were collected retrospectively. The case data were collected through the medical history registered in the manual and the outpatient medical record system. Telephone follow-up was conducted among the patients who had received menopausal hormone therapy to know about whether they were taking medication and to record the reasons for treatment interruption. Univariate analysis and multivariate Logistic regression analysis were adopted to investigate the influencing factors of discontinuing menopausal hormone therapy in patients with menopausal syndrome. RESULTS A total of 183 patients receiving menopause hormone therapy were enrolled. They were divided into interruption group （78 cases） and continuation group （105 cases） according to whether the treatment was interrupted. The results of telephone follow-up showed that the reasons in turn for interruption were perceiving ineffectiveness （16.67%）， concerning about medication risk（15.38%）， the existence of caution case（12.82%） and adverse reactions（10.26%）. The results of univariate analysis showed that there were statistically significant differences in occupation， complications， medication regimen， bone condition and blood viscosity between the two groups （P＜0.05）. Multivariate Logistic regression analysis showed that the absence of complications， osteopenia and osteoporosis， working in public institution and retirement， and the continuous sequential medication regimen favored continuation of menopausal hormone therapy （P＜0.05）. CONCLUSIONS The interruption rate of menopausal hormone therapy is relatively high， and patients are greatly affected by perceiving ineffectiveness and concerning about medication risks， the existence of caution case， and adverse reactions. Complications can cause patients to interrupt treatment， while factors such as osteopenia and osteoporosis， working in public institutions and retirement， and continuous sequential medication regimens make patients more inclined to choose to continue menopausal hormone therapy.