Quantitative and qualitative analysis of common peaks in chemical fingerprint of Yuanhu Zhitong tablet by HPLC-DAD–MS/MS

A quality control (QC) strategy for quantitative and qualitative analysis of “common peaks” in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DAD–MS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C₁₈ column with a gradient elution using a mixture of 0.4‰ ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the “common peaks”. For quantification of “common peaks”, the detection wavelength was set at 254 nm, 270 nm, 280 nm and 345 nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of “common peaks”, 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LC–MS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT.     

结核分枝杆菌rpoB基因和突变检测技术在结核性脑膜炎中的诊断价值

目的 评价结核分枝杆菌rpoB基因和突变(GeneXpertMTB/RIF)检测技术对结核性脑膜炎(tuberculous meningitis,TBM)的诊断价值。方法 回顾性分析2019年1月至2020年6月陕西省结核病防治院收治的同时采用抗酸杆菌(AFB)涂片镜检(简称“AFB涂片”)、BECTECMGIT960液体培养(简称“MGIT960培养”)和GeneXpertMTB/RIF(简称“GeneXpert”)检测的142例疑似TBM患者,其中确诊TBM患者82例、非TBM患者60例。以临床确诊TBM患者为参考标准评价AFB涂片、MGIT960培养和GeneXpert检测对TBM的诊断效能。结果 AFB涂片、MGIT960培养和GeneXpert检测的敏感度分别为3.7%(3/82),24.4%(20/82)和40.2%(33/82);特异度均为100.0%(60/60);符合率分别为44.4%(63/142),56.3%(80/142)和65.5%(93/142)。结论 AFB涂片技术在诊断TBM时的敏感度很低,MGIT960培养检测技术在诊断TBM时的敏感度较高,而GeneXpert检测技术在诊断TBM时的敏感度更高。     

Use of intrathecal midazolam or fentanyl as an adjunct to spinal anaesthesia with bupivacaine for lower limb surgery: A randomised controlled study

BackgroundIntrathecal adjuncts have been used to prolong the quality and duration of subarachnoid anaesthesia. Midazolam given intrathecally is reported to have antinociceptive properties. The purpose of this study was to compare intrathecal midazolam with fentanyl for pain relief and patient comfort.MethodsIn this prospective, double-blind, randomised controlled trial, 90 patients for lower limb surgeries were randomly allocated to three groups: “control group” (3 ml 0.5% heavy bupivacaine), “fentanyl group” (3 ml 0.5% bupivacaine + 10 mcg fentanyl) and “midazolam group” (3 ml 0.5% bupivacaine + 1 mg midazolam). Level, duration, and quality of blocks were compared along with the duration and quality of postoperative analgesia. Haemodynamic stability and any associated complications were also noted.ResultsOnset of block was fastest in Group C (28.5 ± 13.48 min) vis-a-vis other groups (35.5 ± 26.05 min for Group “F” and 28.5 ± 23.68 min for Group “M”; P = 0.51). Duration of block was comparable in all groups (130.5 ± 39.3 min Group “C”; 126.5 ± 44.0 min Group “F” and 129.5 ± 45.7 min Group “M”; P > 0.5).Addition of adjuncts did not significantly defer the appearance of pain. Intensity of pain was lower in Group “M”. Average VAS scores were lower for Group “M” (3–4) than those for Group “C” (4–5) and Group “F” (4–6).Majority of patients required at least one dose of rescue analgesic; however, those receiving fentanyl reported better quality of postoperative analgesia than those in midazolam group.ConclusionAdjuvants improve quality of postoperative analgesia (fentanyl better than midazolam).     

“基于问题学习”在线教学方案的实施效果

目的分析“基于问题学习(problem-based learning,PBL)”在线教学方案的实施效果,为提高PBL在线教学质量提供参考。方法2020年1月至5月,制定“视频会议系统+微信群+图书馆VPN在线资源+PBL网站”PBL在线教学方案(简称PBL在线教学方案),选取北京大学2017级八年制基础医学专业、2017级八年制和五年制临床医学专业共363名学生为研究对象,通过问卷调查和PBL网站反馈,以描述性分析和χ²检验分析PBL在线教学方案的实施效果。结果90.9%(260/286)的学生反馈网络情况“很好”或“还行”。学生对PBL在线教学方案中学生主席、记录员、组员和指导教师的认可度分别为83.6%(239/286)、92.0%(263/286)、64.0%(183/286)和87.1%(249/286)。76.9%(220/286)的学生对讨论效果满意。关于学生对PBL在线教学和线下教学的学习效果反馈,在“案例学习提高对PBL的兴趣”[81.4%(763/937)比83.0%(1698/2047)]和“资讯收集、分析、整理和批判性阅读的提高”[87.8%(823/937)比89.8%(1839/2047)]2个方面,其差异均无统计学意义(均P>0.05);在“明确案例学习目标和重点,并掌握用于解释临床现象的机制”方面[85.4%(800/937)比90.4%(1852/2047)],其差异具有统计学意义(P<0.01)。结论网络情况能够保证PBL在线教学方案的实施;学生对PBL在线讨论时记录员和指导教师的认同度高;PBL在线教学效果基本达到线下教学效果。PBL在线教学方案对PBL混合式教学可以提供一定借鉴。     

A study of the effect of “Strengthening body Resistance Method” on asthma attack

In order to study the effect of a traditional Chinese medical treating method (Strengthening Body Resistance Method) on asthma attack, asthmatic patients of either Cold type or Heat type were divided into two groups and treated with the Chinese methods “Zhibiao” (symptomatic treatments, as the control group) and “Biaoben Jianzhi” (treating both the secondary and principal aspects of a disease, as the test group) respectively. The results showed that the markedly effective rates were higher in the two “Biaoben Jianzhi” groups than that in the two “Zhibiao” groups. After treatment, the changes of one second forced expiratory volume (FEV₁) and the peak expiratory flow of patients of the former were also greater than those of the latter; the proportion of cells carrying human leukocyte antigen class II (HLA-DR⁺ cell), the basophilic cell releasing capacity (HBR) and the T cell hyperplasia response to specific allergens of patients of the “Biaoben Jianzhi” groups were all reduced while those of the “Zhibiao” groups changed little. These results suggested that the “Strengthening Body Resistance method” played an important role in regulating immunity for relieving asthma attack.     

人全血他克莫司浓度高效液相色谱-质谱法检测方法学的建立及性能评价

目的 建立全血他克莫司高效液相色谱-质谱(HPLC-MS)的检测方法.方法 液液萃取的方法从血中提取出他克莫司,然后用高效液相色谱-质谱的方法分离并检测他克莫司的浓度;同时进行标准曲线、线性范围、最低检测限、精密度、回收率及比对实验.结果 方法标准曲线回归方程为Y=0.2464x+0.0082,R2=0.9996;线性范围为0.100～40.000 ng/ml,回归方程为Y=1.0294x-0.035,R2=0.9998;最低检测限为0.100 ng/ml;以浓度为0.300、4.004、8.008、16.016 ng/ml的4种浓度样本测定不精密度(CV),其批内CV分别为4.67%、2.78%、3.22%、3.58%,其总CV分别为5.67%、3.16%、3.97%、4.29%;用浓度为0.300、4.004、8.008、16.016 ng/ml 4种标准品进行回收实验,其回收率分别为95.67%、97.00%、95.88%、98.30%;与酶联免疫吸附试验(ELISA)方法进行比对,回归方程为y=1.0172x+0.3742,相关系数R2=0.9630,2种方法相关性较好,ELISA方法的测定值较HPLC-MS方法的测定值高,差异有统计学意义(P<0.01).结论 HPLC-MS方法具有方便、快速、准确和成本低的优点,可用于对全血中他克莫司的临床监测和实验研究.     

Sensitivity and specificity of an enzyme-linked immunosorbent assay for the detection of infectious bovine rhinotracheitis viral antibody in cattle

An enzyme-linked immunosorbent assay was developed to detect bovine serum antibody to infectious bovine rhinotracheitis virus. The specificity of this assay in 304 bovine sera, collected from an infectious bovine rhinotracheitis virus-free herd, was 100%; in sera from 62 cattle inoculated with an intranasal vaccine, its diagnostic sensitivity was 27.4% at one month and 100% at six months, postvaccination. In 303 bovine sera with standard serum neutralizing antibody titers of greater than or equal to 1:2 it showed 100% sensitivity; and in 463 random diagnostic samples, comparative tests indicated that enzyme-linked immunosorbent assay detected more seropositive animals (61.6%) than the standard serum neutralizing test (49.9%). The enzyme-linked immunosorbent assay method was considered to be technically superior as a routine diagnostic test for the detection of infectious bovine rhinotracheitis viral antibody in bovine sera.     

分光光度法测定中药及制剂成分含量时空白液制备方法研究

建立薄层洗脱方法制备中药及制剂的空白液,考察药典载马钱子、槐米、复方银黄口服液的分光光度测定法,结果表明空白液的干扰占其含量的16%以上.建议用分光光度法测定中药及制剂中成分含量时,应考虑被测药本身所含的干扰成分.     

结核分枝杆菌潜伏感染早期两种检测方法的比较分析

目的比较结核分枝杆菌(Mycobacterium tuberculosis,MTB)特异性IFN-γ酶联免疫斑点法(Elispot)和结核菌素皮试(TB-PPD皮试)在MTB潜伏感染早期诊断的优异。方法采用自主研制的Elispot试剂盒(简称Elispot)对100例健康志愿者进行检测,并和TB-PPD皮试进行平行比较。结果100例健康志愿者中有50例TB-PPD皮试阳性(占50%),Elispot检测阳性20例(占20%)。两种方法检测结果差异有统计学意义(χ2=20.45,P<0.005)。结论MTB特异性IFN-γElispot试剂盒在健康人群中检测阳性率比传统的TB-PPD皮试要低,Elispot检测体外IFN-γ应答反应在诊断MTB潜伏感染的价值优于TB-PPD皮试,在MTB潜伏感染早期诊断有较好的发展前景。     

Roles of Rho/Rock Signaling Pathway in Silica-induced Epithelial-mesenchymal Transition in Human Bronchial Epithelial Cells

ObjectiveTo investigate the roles of Rho/Rock signaling pathway in silica-induced Epithelial-mesenchymal transition (EMT) in human bronchial epithelial cells (BEC) in vitro.MethodsHuman BEC were incubated with silica with various concentrations for indicated times. Cell viability was assayed by MTT test. Morphologic Changes were observed by microscope. Mesenchymal marker α-smooth muscle actin (α-SMA), vimentin (Vim), and epithelial marker E-cadherin (E-cad) were analyzed by Western Blot. The pull-down assay was used to measure Rho activity. In the prevention experiments, the specific inhibitor for Rho effector ROCK (Y27632) was used to inhibit the activity of Rho.ResultsHuman BEC stimulated with silica were converted from a “cobblestone” epithelial structure into an elongated fibroblast-like shape structure. Incubation of human BEC with silica induced de novo expression of α-SMA and Vim, and loss of E-cad. Also, silica treatment resulted in Rho activation in human BEC. Y27632 up-regulated the E-cad expression but attenuated α-SMA and Vim expression in silica-stimulated cells.ConclusionThe activation of Rho/ROCK signaling pathways is most likely involved in Silica-induced EMT in human bronchial epithelial cells.     

昆明小鼠旷场实验评价方法的三维结构

目的 探讨旷场实验(Open Field Test,OFT)作为昆明小鼠行为学评价方法的结构维度.方法成年雄性昆明小鼠放入旷场箱正中央格,摄像系统记录5 min的行为变化,实验间隔1周进行2次;采用因子分析进行分析评价,实验参数如下:中央区停留时间百分率( Ctime％)、中央区水平运动百分率( Ccross％)、水平运动(Cross)、垂直运动(Rear)和粪便粒数(FB).结果 Ctime％与Ccross％ (P=0.756,P ＜0.001:Pearson=0.869,P＜0.001)、Rear(P=0.694,P＜0.01;Pearson=0.465,P＜0.05)及Cross与Rear(P=0.599,P＜0.01:Pearson =0.739,P＜0.001)初测重测均具有较好组内相关性;Ctime％ (P=0.586,P＜0.01)、Ccross％(P=0.559,P＜0.05)、Cross(P=0.633,P＜0.01)、Rear(P=0.612,P＜0.01)初测重测均具有较好相关性;OFT所有参数初测重测可提取三个公因子:Ctime％、Ccross％对焦虑因子F1贡献大(Loading％:43.34％,48.56％),Cross、Rear对运动探索因子F2贡献大(Loading％:27.94％,29.30％),FB对情绪因子F3贡献大(Loading％:21.95％,15.93％);F1、F2((P=0.567,P＜0.01;Pearson =0.538,P＜0.05)初测重测相关性亦较好.结论 OFT作为昆明小鼠行为学评价方法,具有三维结构:焦虑因子维度、OFT活性因子维度(运动探索因子)和情绪因子维度.     

RD105基因缺失法和双重PCR法对125株结核分枝杆菌菌株基因分型检测

目的 为明确比较RD105基因缺失法和双重PCR法对辽宁地区结核分枝杆菌中北京家族菌株的检测能力，同时掌握辽宁地区结核杆菌中北京家族菌株的流行情况而进行该项研究。方法 选取2011—2013年间辽宁省内各地市临床分离菌株，随机抽取经传统方法菌种鉴定为结核分枝杆菌的菌株，共计125株。选用超声破碎法提取菌株DNA，应用RD105基因缺失法和双重PCR法，同时对收集到的125株结核分枝杆菌进行特异性扩增，最后使用琼脂糖凝胶水平电泳对扩增的结果进行判断。结果 两种方法判断标准为：RD105基因缺失法结果中，出现283 bp大小片段的为北京家族菌株，不出现特异性条带的为非北京家族菌株；双重PCR法结果中，只出现393 bp大小片段的为北京家族菌株，而只出现570 bp大小片段为非北京家族菌株。使用上述两种方法鉴定本研究的125株结核分枝杆菌，结果完全相同，其中北京家族菌株有110株（88%），非北京家族菌株有15株（12%）。结论 RD105基因缺失法和双重PCR法对辽宁地区结核分枝杆菌中北京家族菌株的检测能力相当，并且北京家族菌株在辽宁地区流行的结核分枝杆菌中为绝对优势株；两种方法相对于传统的Spoligotyping法更简单、更省时，检测成本也较低，更适合在我国多数实验室推广应用。     

应用反相高效液相色谱法同时测定积雪草提取物中羟基积雪草苷、积雪草苷及其苷元

目的：采用反相高效液相色谱建立同时测定积雪草提取物中羟基积雪草苷、积雪草苷、羟基积雪草酸及积雪草酸4种主要成分的分析方法。方法：采用Agilent 1100 高效液相色谱仪系统,ZORBAX Eclipse XDB-C8色谱柱(4.6 mm×150 mm, 5 μm);流动相为1 mmol/L 磷酸二氢钾水溶液-乙腈,梯度洗脱;检测波长205 nm。结果：羟基积雪草苷、积雪草苷、羟基积雪草酸及积雪草酸的色谱响应线性范围良好（R²≥0.9998）;平均回收率分别为97.4%、93.7%、97.5%、99.8%,相对标准偏差值分别为3.4%、1.4%、4.7%、4.4%（n=9）。结论：采用反相高效液相色谱建立同时测定羟基积雪草苷、积雪草苷及其苷元的方法高效灵敏、重现性好,可以作为积雪草提取物质量控制的检测方法。     

The Validity of Cognitive Testing in Screening for Dementia

The concept of “cognitive impairment,” as an indicator of dementia, defined in 1980 as a loss of intellectual abilities of sufficient severity to interfere with social or occupational functioning, is the model which has been adopted for a condition, which has recently been reclassified from an “organic” to a “cognitive disorder.” Data derived from the assessment of a sample of older people demonstrated the extreme sensitivity of a widely employed cognitive assessment instrument to all levels of educational experience, and educational correlates, notably level of physical disability, were identified as other independent predictors of test performance. The analyses raise questions with regard to the reinterpretation of “lack of education,” from a confounding factor in prevalence estimates of cognitive impairment to a “risk factor” for dementia, and support those who have questioned the validity of the one-dimensional “cognitive paradigm,” and the trend to diagnosis based upon objective assessment with standardized instruments.     

Serum Lipoprotein Concentration and Composition in Healthy 50-year-old Men

The classification of hyperlipoproteinaemias (HLP) is based on arbitrary limits between “normolipidaemia” and “hyperlipidaemia”. 92 randomly selected healthy 50-year-old men were studied regarding serum lipoprotein (LP) lipid composition to define limits for “normality”. Cholesterol and triglyceride concentrations were determined and the individual ratios between cholesterol and triglycerides were calculated in the ultracentrifugally isolated LP density classes. A significant linear correlation between cholesterol and triglyceride concentrations was found in the very low density lipoproteins (VLDL) and low density lipoproteins (LDL) but not in the high density lipoproteins (HDL). When different percentile limits for LDL cholesterol and VLDL triglycerides were used as cut-off points for “normality” not only the absolute prevalence but also the relative frequency of different types of HLP was influenced. 26 overweight men studied separately showed significantly increased VLDL cholesterol and triglyceride levels with maintenance of the normal ratio cholesterol/triglycerides of this LP class compared with non-obese subjects. Three different kinds of “extra bands” were occasionally seen on agarose electrophoresis: a “double pre-beta” band in whole serum (3%), a “sinking pre-beta” band with density >1.006 (17%) and a “late pre-beta” band in VLDL (22%). While “sinking pre-beta” is identical with Lp (a) LP the “late pre-beta” band probably represents a certain accumulation of “intermediary particles”.     

电针对自体髓核移植大鼠脊髓后角小胶质细胞活性的影响?

目的观察近部取穴、远部取穴、远近配穴电针对自体髓核移植大鼠模型痛行为及脊髓后角小胶质细胞标记物OX42和环氧化酶2(COX2)表达的影响,为理解不同针灸处方疗效差异提供依据。方法雄性SD大鼠54只,随机分为正常组、模型组、假手术组、远近配穴组、近穴组、远穴组,每组9只。建立自体髓核移植疼痛大鼠模型,远近配穴组取双侧"腰5夹脊穴""大肠俞""委中"和"昆仑",近穴组取双侧"腰5夹脊穴"和"大肠俞",远穴组取双侧"委中"和"昆仑",每天1次,每次20 min,共治疗7 d。测定大鼠热刺激爪退缩阈值(TWL),用免疫组织化学法检测大鼠脊髓后角左侧OX42及COX2的表达变化。结果自体髓核移植大鼠TWL上升,脊髓后角OX42和COX2表达增加。与模型组相比,治疗组可以改善TWL(P0.05),抑制脊髓后角OX42表达,显著抑制脊髓后角COX2表达(P0.05),远近配穴和近部取穴的抑制作用优于远部取穴。结论无论是近部取穴、远部取穴和远近配穴电针均能抑制脊髓后角炎性信号通路发挥抗炎效应,远近配穴和近部取穴的疗效优于远部取穴。     

关于《伤寒论》中药物用量规律的分析

目的根据易学的象数关系理论探讨《伤寒论》中用药与功效的关系。方法采用《伤寒论》原文中的剂量用数和后人对每个药物在不同方中功效的分析,应用Logistic回归、配对卡方检验等统计方法进行量与效关系的研究。结果药物剂量用数与汗、吐、下、和、温、消、清、补8种功效虽有相关,但不能用数来完全解释功效。结论探讨出一种研究《伤寒论》用药规律的方法 ,为后续研究提供思路。     

Quantitative assessment of prevalence of pre-analytical variables and their effect on coagulation assay. Can intervention improve patient safety?

BackgroundVery few Indian studies exist on evaluation of pre-analytical variables affecting “Prothrombin Time” the commonest coagulation assay performed. The study was performed in an Indian tertiary care setting with an aim to assess quantitatively the prevalence of pre-analytical variables and their effects on the results (patient safety), for Prothrombin time test. The study also evaluated their effects on the result and whether intervention, did correct the results.MethodsThe firstly evaluated the prevalence for various pre-analytical variables detected in samples sent for Prothrombin Time testing. These samples with the detected variables wherever possible were tested and result noted. The samples from the same patients were repeated and retested ensuring that no pre-analytical variable is present. The results were again noted to check for difference the intervention produced.ResultsThe study evaluated 9989 samples received for PT/INR over a period of 18 months. The prevalence of different pre-analytical variables was found to be 862 (8.63%). The proportion of various pre-analytical variables detected were haemolysed samples 515 (5.16%), over filled vacutainers 62 (0.62%), under filled vacutainers 39 (0.39%), low values 205 (2.05%), clotted samples 11 (0.11%), wrong labeling 4 (0.04%), wrong vacutainer use 2 (0.02%), chylous samples 7 (0.07%) and samples with more than one variable 17 (0.17%). The comparison of percentage of samples showing errors were noted for the first variables since they could be tested with and without the variable in place. The reduction in error percentage was 91.5%, 69.2%, 81.5% and 95.4% post intervention for haemolysed, overfilled, under filled and samples collected with excess pressure at phlebotomy respectively.ConclusionCorrecting the variables did reduce the error percentage to a great extent in these four variables and hence the variables are found to affect “Prothrombin Time” testing and can hamper patient safety.     

血必净注射液对58例老年脓毒症高热患者体温控制的临床观察

着重对血必净注射液对老年脓毒症高热患者体温的控制情况作一临床观察,其结果表明血必净在老年脓毒症的辅助治疗中对患者体温的控制有明显作用且这种良好的控制作用与其应用时间有一定的相关性。     

“品管圈”活动在降低新生儿留置胃管非计划性拔管中的应用效果分析

目的：探讨“品管圈”活动在降低新生儿留置胃管非计划性拔管中的应用效果。方法回顾分析2013年1月～2013年4月成立“品管圈”活动前190例患儿留置胃管非计划性拔管情况及2013年5月～2013年12月成立“品管圈”活动后243例患儿留置胃管非计划性拔管情况,进行现状把握,目标设定,原因解析,开展对策拟定实施及检讨,效果确认,制定留置胃管固定的标准化流程。结果新生儿留置胃管非计划性拔管率由活动前的74%降至活动后的32%,差异有统计学意义（χ2=19.12,P＜0.05）。结论“品管圈”活动能有效降低新生儿留置胃管非计划性拔管率,提高了护士护理科研参与意识及护理工作主动性和创造性。