基于实时阻抗细胞分析技术评价H9c2心肌细胞缺氧/复氧损伤模型及三七皂苷R_1的干预作用

目的基于实时阻抗细胞分析技术,建立规范标准的H9c2心肌细胞缺氧/复氧损伤模型,并用三七皂苷R_1进行评价。方法将H9c2心肌细胞接种到E-Plate板中,设3个复孔,分别测定生长曲线、不同缺氧时间、不同复氧时间及三七皂苷R_1的干预下细胞指数值,以细胞指数值反映H9c2心肌细胞的生长状态。结果本研究发现,缺氧前更换无血清、无糖DMEM培养基,缺氧4 h,复氧16 h,细胞存活率为(48.82±5.32)%,与MTT法测定结果差异无显著性,且三七皂苷R_1能够保护缺氧/复氧处理H9c2心肌细胞损伤,不存在时间依赖性。结论实时阻抗细胞分析技术能够建立标准规范的缺氧/复氧模型方法,对发现新药物靶点及作用机制也有一定的指导意义。

Study on relationship between perifollicular blood flow and in vitro fertilization-embryo transfer

Objective：To study the relationship between perifollicular blood flow and follicule development, oocyte maturing rate, fertilizing rate, cleaving rate, embryo quality and the outcomes of embryo transfer. Methods： The samples were selected from 66 suffers who underwent in vitro fertilization（IVF）or intracytoplasmic sperm injection（ICSl）. Eeach patients＇ perifollicular blood flow（diameter ≥ 12mm ）was estimated on the day of human chorionic gonadotropin（HCG）administration. Results：Among 66 cycles, 26（39.4%） cycles resulted in pregnancy, perifollicular blood flow resistance index（Rl）, peak systolic velocity/end diastasis velocity（S/D） of non-pregnant group was significantly higher than that of the pregnant group （P 〈 0.004）. When RI 〈 0.49, the pregnancy rates, fecundation rates, fertilization rates, metaphase numbers for the of second meiosis oocytes increased evidently （P 〈 0.05）, but there were no statistical difference in gonadotropin dosage, cycle frequency, infertility years, ages, estradiol（E2）on the day of HCG administration, numbers of oocyet retrieved and high-quality embryo rates （P 〉 0.05 ）. There were no statistical difference between non-pregnant group and pregnant group in S and D （P 〉 0.05 ）. There was no correlation between periFollicular blood flow RI and follicular diameter by linear regression analysis. Conclusion：Our study shows that perifollicular blood flow RI and S/D are effective indices of predicting the pregnancy outcome of IVF-ET.

基于指纹图谱分析和多成分同时定量的益安宁丸质量评价研究

目的:建立益安宁丸的HPLC指纹图谱,并测定5种主要成分(人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1、五味子酯甲和丹参酮ⅡA)的含量,为益安宁丸的质量控制提供可靠方法。方法:采用Waters Sunfire TM-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱(0~5 min,2%A;5~15 min,2%A→10%A;15~25 min,10%A→30%A;25~40 min,30%A;40~60 min,30%A→90%A;60~70 min,90%A);流速为1.0 ml·min^-1;检测波长为203 min(0~60min,检测人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1和五味子酯甲)、270 nm(60~70 min,检测丹参酮ⅡA);柱温35℃。结果:在指纹图谱研究中,共确定益安宁丸HPLC指纹图谱26个共有峰,通过与混合对照品比较指认其中7个指标成分,分别是6号-五味子酯甲、16号-人参皂苷Rg1、17号-人参皂苷Rb1、18号-三七皂苷R1、23号-丹参酮Ⅰ、24号-五味子甲素和26号-丹参酮ⅡA,利用相似度软件对14批样品指纹图谱进行分析,各批样品相似度均在0.98以上。在建立的色谱条件下试验5个成分分离度良好,方法精密度和重复性RSD值均<1.5%,供试品溶液在24 h内稳定,各成分具有良好的线性关系和较宽线性范围;6批益安宁丸中人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1、五味子酯甲和丹参酮ⅡA质量分数分别在7.1870~7.2540,9.0860~9.2500,2.1090~2.2880,11.2080~11.3380和1.3740~1.4580 mg·g^-1。结论:所建立的益安宁丸HPLC指纹图谱检测和定量测定分析方法快速、准确,可以有效地评价益安宁丸的质量。