Suitability of a liquid chromatography assay of neomycin sulfate to replace the microbiological assay for neomycin in USP Monographs

The current USP National Formulary contains 65 Monographs for drug formulations containing neomycin. All 65 Monographs prescribe a bioassay for neomycin assay. This bioassay, based on cell culture, is labor intensive, has poor precision, and cannot be adapted for purity or identification. High-performance anion-exchange chromatography with integrated pulsed amperometric detection (HPAE-IPAD), a liquid chromatography technique, has been shown to be suitable for neomycin purity analysis and neomycin assay of an over-the-counter first aid cream (Hanko and Rohrer [17]). Here we propose that an HPAE-IPAD assay can replace the bioassay in the 65 neomycin-containing Monographs. We applied the HPAE-IPAD assay to four neomycin-containing drug products representing the four classes of formulations found in the 65 Monographs, liquid, solid, suspension, and cream. Each drug was analyzed with two chromatography systems, and on 3 separate days. For all products, HPAE-IPAD measurements were precise and accurate with respect to the label concentrations. There was also high accuracy for spike recovery of neomycin from the four drug products throughout 70–150% of the labeled concentration. These results suggest that an HPAE-IPAD assay would be an accurate assay for neomycin, and would be faster and more precise than the current bioassay.     

从COS搁置谈原料药生产企业如何通过国际认证现场检查

目的:为我国原料药出口生产企业顺利通过国际认证现场检查提供借鉴。方法:跟踪国外原料药进出口相关法规变化,分析我国原料药生产企业欧洲药典适用性证书(COS)被搁置的原因并提出建议。结果与结论:生产现场存在与欧盟《药品生产质量管理规范》(GMP)要求不符的重大缺陷是导致COS被搁置的主要原因,提高原料药生产过程的GMP管理水平是企业顺利通过国际认证现场检查的关键所在。     

单孔悬吊与气腹腹腔镜在子宫肌瘤切除术中的比较

目的 探讨单孔悬吊式腹腔镜在子宫肌瘤切除术中的应用价值.方法对比分析2010年10月～2011年10月开展单孔悬吊与气腹腹腔镜子宫肌瘤切除术各45例的手术情况.结果 单孔悬吊式腹腔镜组在手术时间、术中出血量、术后排气时间、术后并发症及平均住院费用方面均优于气腹组.结论 单孔悬吊式腹腔镜切除肌瘤,拓宽了气腹腹腔镜手术范围,降低了手术风险及设备投资,减少了术后并发症及住院费用,在妇科手术中值得推广.     

Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)

Efficient vaccine production requires the growth of large quantities of virus produced with high yield from a safe host system. Human influenza vaccines are produced in embryonated chicken eggs. However, over the last decade many efforts have allowed the establishment of cell culture-derived vaccines.We generated a Vero cell line adapted to grow in suspension (sVero) in a serum-free medium and evaluated it for its potential as host cell for influenza vaccine production. Initially we studied the capacity of sVero cells to grow in the presence of incremental concentrations of trypsin. In comparison with adherent Vero cells (aVero), we found that sVero cells maintain their growth kinetics even with a three-fold increase in trypsin concentration.The influence of the conditions of infection on the yield of H1N1 produced in serum-free suspension cultures of sVero cells was investigated by a 2² full factorial experiment with center point. Each experiment tested the influence of the multiplicity of infection (m.o.i.) and trypsin concentration, on production yields at two levels, in four possible combinations of levels and conditions, plus a further combination in which each condition was set in the middle of its extreme levels.On the basis of software analysis, a combination of m.o.i. of 0.0066 TCID_50%/cell and trypsin concentration of 5 μg/1.0 × 10⁶ cells with a desirability of 0.737 was selected as the optimized condition for H1N1 production in sVero cells.Our results show the importance of proper selection of infection conditions for H1N1 production on sVero cells in serum-free medium.     

Laparoendoscopic Single-site Surgery for Comprehensive Staging of Early Ovarian Cancer

Study ObjectiveTo demonstrate the procedure and suspension skills of laparoendoscopic single-site (LESS) staging surgery with infrarenal para-aortic lymphadenectomy for early-stage ovarian cancer.DesignA presentation of the surgery through this technical video.SettingA hospital.Patient and InterventionsA 45-year-old woman presented with a pelvic mass on gynecologic examination and a serum cancer antigen 125 level of 5910 U/mL (normal = <35 U/mL). A computed tomographic scan revealed a mixture of solid and cystic components (70 × 77 × 71 mm) arising from the right ovary and characterized by the “ovarian vascular pelvic” sign. Clinically early-stage ovarian cancer was suspected. Subsequently, LESS staging surgery was performed by an experienced surgeon in our department.ResultsThe surgery lasted 280 minutes, and the volume of blood loss was 50 mL; there were no intra- or postoperative complications. We “hid” the incision perfectly for cosmetic purposes. The histopathologic findings supported high-grade serous ovarian cancer of the right ovary with the left fallopian tube involved as well. In addition, a total of 34 negative pelvic and 18 negative para-aortic lymph nodes were identified, and a stage of IIA was diagnosed as a result.ConclusionWe performed an LESS staging surgery for early-stage ovarian cancer successfully. Our video shows that the LESS approach provided feasible, cosmetic, and safe access among the selected malignant gynecologic surgery. Therefore, we have experienced that the effective suspension was an auxiliary measure for LESS lymphadenectomy. In addition, compared with multiport laparoscopy, the LESS approach could provide easier access to infrarenal para-aortic regions; furthermore, it was safe and quick to extract an unknown sample.     

膝髋关节置换术非计划性暂停手术的原因分析

目的：分析择期膝、髋关节置换术前非计划性暂停手术的现状和原因,寻找其中的规律和共同性,为进一步优化择期膝、髋关节置换的患者术前管理,提高医疗资源的使用效率提供依据。方法： 选择北京大学第三医院骨科2013年9月至2014年8月住院拟择期行膝、髋关节置换手术,但术前因故被暂停手术的病例资料,记录这些病例被暂停手术的原因,对其进行分析。结果： 共拟行膝、髋关节置换手术1 146台,其中完成1 003台,因故暂停143台,停手术率达12.5%。暂停手术最常见的原因是心血管系统疾病（31%）,其他部位感染（14%）, 菌尿（13%）和手术适应证不强（11%）,其他导致手术暂停的原因还有血液系统检查异常、下肢静脉血栓、肝功能异常、血糖控制欠佳等。在不同性别的患者之间,停手术率差异无统计学意义（男性 15.0%,女性11.6%, P=0.149）,不同年龄段的患者之间,停手术率差异无统计学意义（≤50岁 13.0%, 51~65岁 11.6%,66~80岁 13.3%,>80岁11.1%,P=0.864）,但髋关节置换较膝关节置换更易被暂停手术（膝关节置换11.1%,髋关节置换 16.1%,P=0.021）。结论： 对于择期行膝、髋关节置换的患者进行恰当的术前教育和管理非常重要,通过明确诊断、细致的病史询问和体格检查,以及有针对性的检查化验,尽量避免住院后暂停手术的发生,提高医疗资源的使用效率。     

细胞密度对卵巢癌干细胞悬浮球诱导的影响

目的 研究细胞密度是否会对从人卵巢透明细胞癌系ES2 和卵巢腺癌细胞A2780中分离的癌干细胞产生影响。方法 将ES2、A2780细胞置于含生长因子、牛血清白蛋白和人胰岛素的无血清培养基中培养,采用连续传代诱导培养癌干细胞。观察不同细胞密度下ES2、 A2780细胞形态变化,流式细胞术分别检测含血清培养基（SSM）组、无血清培养基（SFM）组细胞诱导前后细胞表面标志CD133、CD44表达的改变。软琼脂克隆形成实验检测不同诱导条件下ES2癌干细胞球生长速度和成瘤能力。结果 2×104 mL-1密度下,ES2细胞能在SFM中存活,但不能形成癌干细胞球;5×104、1×104 mL-1 ES2细胞能在SFM中存活、增殖并形成癌干细胞球。与贴壁的细胞及诱导前相比,诱导后形成的癌干细胞球（1×105 mL-1组、5×105 mL-1组）高表达CD133+、CD44+ （ P<0.05）,传代后具有更高的增殖能力和克隆形成能力,且5×104 mL-1密度下,癌干细胞球成瘤能力更强。A2780细胞分别以1×104、3×104、5×104 mL-1的密度接种于SFM中,10 d后,1×104、3×104 mL-1组中形成细胞团,但3×104 mL-1组中形成的细胞团更大,透光性更强,细胞更致密,5×104 mL-1组未形成细胞悬浮球。流式细胞术发现A2780细胞3×104 mL-1组中细胞与1×104 mL-1组相比,CD133+细胞阳性率更高,差异有统计学意义( P<0.05)。3×104 mL-1组的细胞生长速度最快。结论 细胞密度5×104 mL-1和3×104 mL-1分别是诱导ES2、 A2780细胞株卵巢肿瘤干细胞球的合适条件。     

悬灸不同状态犊鼻穴的脑电功率谱密度特征研究

目的:通过提炼热敏态腧穴的脑电功率谱密度特征指标,为临床建立热敏态腧穴的脑电客观检测技术奠定科学基础。方法:选取2016年5月至2018年12月江西中医药大学附属医院收治的膝关节骨性关节炎患者30例作为研究对象,以犊鼻穴为试验腧穴,以同侧犊鼻穴正上4寸为对照点,采用热敏腧穴灸感法进行检测,根据施灸过程中是否发生热敏现象分为热敏态腧穴组和非热敏态腧穴组。受试者脑电数据采用美国EGI公司生产的128导高密度脑电系统采集,其采集分为3个时段,每个时段记录15 min,共记录45 min:第1段在艾灸前记录15 min;第2段在艾灸15 min后开始记录15 min;第3段在艾灸结束后开始记录15 min。脑电信号的预处理采用Matlab2014a(美国The MathWorks公司)数据分析软件及其EEGLAB子软件,对灸前、灸中、灸后平均脑电功率谱密度分别进行提取,所得脑电数据采用SPSS 19. 0软件进行统计学分析比较。结果:热敏态与非热敏态腧穴功率谱密度比较:平均功率谱密度差异:热敏态腧穴灸中、灸后与灸前比较,全部导联叠加后的脑电平均功率谱密度有明显增加。进一步分析不同频段脑电功率谱密度分布差异,其中theta、alpha1、alpha 2及beta频段均增加,尤其以alpha1、alpha 2频段增加显著,且beta频段的功率谱密度持续增加至灸后阶段。热敏腧穴不同脑区平均功率谱密度结果提示,灸中、灸后与灸前比较,灸中、灸后双侧额区、顶区、枕区平均功率密度明显增加,而其中央区差异无统计学意义。结论:alpha1、beta节律的能量在热敏态腧穴被激活时占整个脑电能量中的比重较大,而非热敏态腧穴脑电信号变化较少,这可能作为腧穴热敏现象的客观显示标签之一。     

《中国药典》2015年版盐酸头孢他美酯干混悬剂有关物质检查方法的商榷

目的：通过对2个厂家8批盐酸头孢他美酯干混悬剂的检验和进一步研究,对该品种有关物质项检查方法进行优化改进。方法：依据《中国药典》2015年版标准进行检验,延长检测时间并考察2个厂家辅料的干扰情况。结果：盐酸头孢他美酯干混悬剂标准中有关物质检查项,需要增加检测时间至主峰保留时间的5倍,对于辅料峰的扣除必要时应采用辅料对照实验进行。结论：建议修订盐酸头孢他美酯干混悬剂标准的有关物质项目,以利于更好地控制该药品质量。     

结膜上穹窿联合筋膜鞘（CFS）悬吊术对治疗严重上睑下垂的意义

目的：为有效治疗上睑下垂患者,临床探究结膜上穹窿联合筋膜鞘悬吊术治疗效果情况。方法随机选取该院2012年1月—2013年1月期间80例因严重上睑下垂入院治疗患者,按入院时间分为对照组40例、观察组40例。对照组采用额肌瓣悬吊术,观察组行结膜上穹窿联合筋膜鞘悬吊术,观察两组治疗效果,同时对患者术后不良反应以及3年期间疾病复发情况进行比较。结果观察组疾病改善有效率为95.0%较对照组90.0%比较差别无意义,两组手术治疗均有良好效果,P>0.05;术后观察组不良反应率、疾病复发率分别为7.5%、2.5%,对照组依次为17.5%、12.5%,对照组患者术后不良反应发生以及疾病再次复发较观察组多,差异有统计学意义P<0.05。结论临床对严重上睑下垂患者使用结膜上穹窿联合筋膜鞘悬吊术,患者上睑能力恢复良好,且术后复发少、不良反应少。