HPLC波长切换法同时测定杜仲双降袋泡剂中7种成分的含量

目的建立高效液相(HPLC)波长切换法同时测定杜仲双降袋泡剂中桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁及槲皮素7种成分含量的方法。方法以80%甲醇为溶剂,加热回流提取;色谱柱:Hydrosphere C18(4.6 mm×200 mm,5μm);柱温:30℃;流动相:乙腈-0.1%磷酸溶液,梯度洗脱;检测波长:208 nm(桃叶珊瑚苷)、235 nm(京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷)、270 nm(芦丁、槲皮素);流速:0.9 ml/min。结果桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁、槲皮素的质量浓度分别在10.97~274.25μg/ml(r=0.999 7)、9.78~244.50μg/ml(r=0.999 8)、6.86~171.50μg/ml(r=0.999 6)、2.47~61.75μg/ml(r=0.999 7)、8.11~202.75μg/ml(r=0.999 1)、4.59~114.75μg/ml(r=0.999 9)、1.85~46.25μg/ml(r=0.999 5)范围内线性关系良好;平均加样回收率分别为99.85%、98.92%、97.52%、97.08%、98.51%、97.10%、96.91%,RSD分别为0.93%、0.74%、1.26%、1.37%、0.88%、1.05%、1.33%。结论所建立的HPLC波长切换法可以同时测定杜仲双降袋泡剂中桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁和槲皮素的含量,方法简便准确、灵敏度高,可用于杜仲双降袋泡剂的质量控制。     

基于限定日剂量和ABC-改进VEN法的2018年亳州市人民医院质子泵抑制剂应用分析

目的:了解亳州市人民医院(以下简称“我院”)质子泵抑制剂(proton pump inhibitor,PPI)的使用情况,明确需重点管控的PPI品规,向临床提供关于PPI选择的相关建议。方法:调查2018年我院PPI消耗数据,采用限定日剂量(defined daily dose,DDD)法和ABC-改进VEN法进行统计分析,并根据分析结果和管理优先级别确定Ⅰ(高)、Ⅱ(低)和Ⅲ(中)组。结果:ABC-改进VEN法基于减轻患者负担和医院成本对PPI进行分类,便于管理。涉及的PPI共18个品规,销售金额合计14051911.21元;其中Ⅰ组包含AN类和DDD分析加入的品规,共5个,销售金额为9405764.53元(占66.94%),代表药品包括注射用泮托拉唑钠(40 mg)、注射用雷贝拉唑钠(20 mg);Ⅱ组包含CV类5个品规,销售金额为886138.62元(占6.31%),代表药品为奥美拉唑肠溶片(20 mg);Ⅲ组包含10个品规,销售金额为3760008.06元(占26.76%),代表药品为奥美拉唑肠溶片(40 mg)。结论:我院PPI应用结构不甚合理,应重点监控Ⅰ组药品,减少不合理用药情况,向临床推荐Ⅲ组药品,并保障供应。     

基于一测多评法对丁蔻理中丸中6种成分的质量控制研究

目的:建立一测多评法同时测定丁蔻理中丸中白术内酯Ⅲ、白术内酯Ⅰ、丁香酚、6-姜辣素、8-姜酚、10-姜酚6种成分的含量。方法:以白术内酯Ⅰ为内标物,建立白术内酯Ⅰ与白术内酯Ⅲ、丁香酚、6-姜辣素、8-姜酚、10-姜酚之间的校正因子,计算待测成分含量,并将外标法测定值与一测多评法计算值进行对比,验证所建立一测多评法的准确性。结果:白术内酯Ⅲ、白术内酯Ⅰ、丁香酚、6-姜辣素、8-姜酚和10-姜酚分别在1.09~21.80μg·mL-1、0.71~14.20μg·mL-1、13.67~273.40μg·mL-1、6.04~120.80μg·mL-1、1.46~29.20μg·mL-1、1.78~35.60μg·mL-1范围内线性关系良好;各成分一测多评法计算值与外标法实测值无显著性差异。结论:利用校正因子对丁蔻理中丸中6种成分的含量测定是可行的,一测多评法可以用于丁蔻理中丸的质量评价研究。     

Comparison of seven different reagents of peroxidase method for small and dense low density lipoprotein cholesterol (sdLDL‐C) measurement

BackgroundWe validated the performance of seven different reagents of peroxidase method for sdLDL‐C in two automatic analyzers that are common in Chinese laboratories.MethodsSeven commercially available sdLDL‐C assays were analyzed with the Beckman AU5400 and Mindray BS2000 automatic analyzers. A total of 336 blood samples were collected and the reference interval was also validated in 298 apparently healthy individuals. Serum samples were used for method comparison of precision, recovery, lower limit of detection, comparison and concurrence analysis, as well as reference interval for the Mindray reagent.ResultsThe repeatability CV% of the seven sdLDL‐C assays were 0.81%~3.66% for Mindray BS2000 and 0.76%~3.91% for Beckman AU5400, while Total CVs for Mindray BS2000 sdLDL‐C assay were 1.34%~4.81%, and that of Beckman AU5400 were 2.25%~10.33%. The measured recovery rates of sdLDL‐C assays were within the allowable ±10% deviation range. There was no obvious difference between the reagents in the lower limit detection. There was a difference between the validation results of the reference range and the manufacturer''s.BSBE, Mindray, and Dongou had a high degree of association with DENKA SEIKEN on Mindray BS2000, while BSBE, Mindray, Dongou and Merit Choice had a high degree of association with DENKA SEIKEN on Beckman AU5400. Passing–Bablok regression showed excellent linear correlation between BSBE and Mindray and DENKA SEIKEN and on Beckman AU5400.ConclusionsOur results indicate that the basic performance can meet the testing requirements, but the comparability between them is still insufficient.     

PKA含量动力学检测方法的研究与建立

目的建立激肽释放酶原激活剂(prekallikrein activator, PKA)含量的动力学检测方法,并对该方法进行验证。方法对比不同样品稀释缓冲液的pH、离子强度,各步骤的孵育时间、孵育温度等因素,确定检测方法的最适条件。并对该方法的准确度、专属性、精密度、线性、稳定性和耐用性进行方法学验证。结果确定以0.05 mol/L Tris-HCl缓冲液(pH8.5、含0.15 mol/L NaCl)稀释样品,在37℃下与激肽释放酶原(PK)试剂混合孵育20 min后加入底物S-2302,检测反应前10 min内吸光度变化率△A405/min。方法验证结果表明PKA含量在(0.5~4.0)IU/mL范围内,△A405/min与含量呈良好的线性关系,标准曲线相关系数R2>0.99,校正标样回收率在96.9%~103.7%。专属性验证表明人血白蛋白和静注人免疫球蛋白(pH4)的辅料、低pH及蛋白质含量等因素对该方法检测PKA含量均无明显影响,各溶液的加标回收率分别为98.0%(0.9%NaCl溶液)、95.3%(0.46%辛酸钠溶液)、96.7%(10%麦芽糖溶液,pH4.0)、94.0%(20%BSA)、94.0%(5%BSA,pH4.0)。在(0.5~4.0)IU/mL内,该方法准确度和精密度均符合要求。其中质控品批内回收率在96.4%~109.5%, CV在0.2%~6.9%,批间回收率在101.5%~102.9%, CV在2.6%~5.9%。PK试剂和S-2302在室温放置6 h内,线性、准确度和精密度均能符合要求,质控品回收率在94.9%~109.9%。采用新建的动力学法和《中国药典》(ChP)2015年版终点法检测本公司生产的20批次人血白蛋白样品中PKA含量的结果对比表明2种分析方法差异无统计学意义(P>0.05),2种方法的相关系数为0.99805(n=20)。结论建立的PKA含量动力学检测方法,具有良好的线性、专属性、准确度、精密度、稳定性、耐用性,相比《中国药典》方法,新建方法更节省分析时间,更为方便、准确、快速,可实现快速测定人血白蛋白和静注人免疫球蛋白(pH4)PKA含量测定。     

Application of Pop-off Method to Bone Marrow and Peripheral Blood Specimens for Purposes of Electron Microscopy

In this paper, we describe a pop-off method applicable to the hematological field. Bone marrow or peripheral blood specimens from patients were placed on a clean glass slide and fixed immediately in 2% glutaraldehyde solution for 10 minutes. For the DAB reaction, the slide was immersed in the DAB reagent for 30 minutes, and post fixed with 1 % OsO₄ solution for 1 hour. Specimens on the slide were washed with buffer solution, dehydrated and polymerized directly on the slide. A gelatin capsule filled with Epon mixed monomer was then reversed over the specimen. After polymerization was completed, the specimen was popped off from the slide to the capsule and trimmed carefully to prepare for ultrathin sectioning. This method allows the entire sequence of tissue preparation to be carried out on the slide, from fixation to embedding, and even, especially in scanty specimens, including the DAB reaction. Electron microscopic findings in specimens prepared by this technique show excellent preservation and the absence of specific artifacts.     

Variants of autophagy-related gene 5 are associated with neuromyelitis optica in the Southern Han Chinese population

Using animal models for autoimmune diseases, we have previously shown that allogeneic bone marrow transplantation (allo BMT) can be used to treat autoimmune diseases. Using cynomolgus monkeys, we have recently developed new BMT methods for the treatment of autoimmune diseases. The methods include the perfusion method (PM) for the collection of bone marrow cells (BMCs), and intra-bone marrow (IBM)-BMT for the direct injection of collected whole BMCs into the bone marrow cavity. The PM, in comparison with the conventional aspiration method, can minimize the contamination of BMCs with T cells from the peripheral blood. Therefore, without removing T cells, no graft-versus-host disease (GvHD) develops in the case of the PM. Since BMCs collected by the PM contain not only hemopoietic stem cells (HSCs) but also mesenchymal stem cells (MSCs), the injection of both cells directly into the bone marrow cavity (IBM–BMT) facilitates the engraftment of donor hemopoietic cells. In organ allografts with IBM–BMT, no graft failure occurs even if the radiation dose is reduced. In addition, IBM–BMT is applicable to regeneration therapy and various age-associated diseases such as osteoporosis, since it can efficiently recruit donor-derived normal MSCs.

We have also found that IBM–BMT in conjunction with donor lymphocyte infusion can prevent GvHD, but suppress tumor growth. We believe that this strategy heralds a revolution in the field of transplantation (BMT and organ allografts) and regeneration therapy.     

FRAN and RBF-PSO as two components of a hyper framework to recognize protein folds

In this paper, an intelligent hyper framework is proposed to recognize protein folds from its amino acid sequence which is a fundamental problem in bioinformatics. This framework includes some statistical and intelligent algorithms for proteins classification. The main components of the proposed framework are the Fuzzy Resource-Allocating Network (FRAN) and the Radial Bases Function based on Particle Swarm Optimization (RBF-PSO). FRAN applies a dynamic method to tune up the RBF network parameters. Due to the patterns complexity captured in protein dataset, FRAN classifies the proteins under fuzzy conditions. Also, RBF-PSO applies PSO to tune up the RBF classifier. Experimental results demonstrate that FRAN improves prediction accuracy up to 51% and achieves acceptable multi-class results for protein fold prediction. Although RBF-PSO provides reasonable results for protein fold recognition up to 48%, it is weaker than FRAN in some cases. However the proposed hyper framework provides an opportunity to use a great range of intelligent methods and can learn from previous experiences. Thus it can avoid the weakness of some intelligent methods in terms of memory, computational time and static structure. Furthermore, the performance of this system can be enhanced throughout the system life-cycle.     

海南文昌城市汉族成人Heath-Carter法体型观察

目的 探讨海南城市汉族体型特征。方法 采用Heath-Carter 人体测量体型方法,对海南文昌城市汉族成年人315例（男为150例,女为165例）进行体型评定。结果 1.海南城市汉族平均体型男性为内胚层-中胚层均衡体型（4.9-5.3-2.0）,女性为偏中胚层的内胚层体型（5.5-4.6-1.9）。2.随着年龄的增长,男性内因子值、中因子值显著增长,外因子值下降,尤以30岁前后这种变化最突出。女性体型30~39岁时较20~29岁身体的充实度有所提高,40~49岁以后则更加结实、丰满。3.男女间体型除20~29、30~39岁组存在显著性差异外,40~49岁以后男女体型差异不显著。4.与其他城市族群比较,海南城市汉族男女平均体型点与山东汉族最接近（男性SAD=0.60,女性SAD=0.40）。结论 海南城市汉族体型接近于中国北方族群。