人巨细胞病毒核酸定量检测试剂盒临床应用及初步评价

目的:探讨人巨细胞病毒(HCMV)核酸定量检测试剂(PCR-荧光探针法)的临床使用及初步评价。方法:用浓缩一步法试剂与煮沸法试剂对192例临床收集的样本进行HCMV DNA检测并对比其结果,同时,将两种试剂的检测结果分别与HCMV-pp65抗原检测结果进行对比。结果:新型人巨细胞病毒核酸检测试剂与煮沸法试剂的阳性一致性百分比98.06%,阴性一致性百分比95.51%,总一致性百分比96.88%;对6例不符样本进行测序复核,并对复核后的结果进行Kappa检验一致性分析,结果 Kappa值=1.000,表明新型试剂的检测结果与煮沸法试剂及复核检测的结果具有很好的一致性。在与HCMV-pp5抗原检测结果的对比中,两种试剂均表现出较好的准确性。结论:人巨细胞病毒核酸检测试剂与煮沸法试剂检测结果具有很好一致性,操作简单快速,减少污染环节,并具有内标控制假阴性,结果准确,符合临床检测要求,具有较高的应用价值。     

自体胸腔积液上清作为培养液对肿瘤浸润淋巴细胞生长的影响

目的：观察自体胸腔积液上清作为培养液对肿瘤浸润淋巴细胞(TIL)生长的影响．方法：恶性胸腔积液中的TIL经贴壁法分离后，分别采用含有细胞因子白细胞介素-2(IL-2)、CD3单克隆抗体(OKT3)及植物血球凝集素(PHA)的自体胸腔积液上清及含10％人AB型血清的RPMIl640进行培养，比较两种培养液对TIL的增殖速度、培养前后免疫表型变化和对自体肿瘤细胞的杀伤活性。结果：培养2周，经统计学比较发现，自体胸腔积液上清作为培养液对TIL的生长速度、免疫表型和对自体肿瘤细胞杀伤活性的影响与应用含10％人AB型血清的RPMIl640培养液培养无差异。结论：自体胸腔积液上清作为培养液培养TIL是可行的。为自体TIL疗法治疗恶性胸腔积液提供理论基础。     

参麦注射液含量测定方法的改进

目的:建立高效液相色谱法测定参麦注射液中人参皂苷Re含量的方法。方法:色谱柱:KromasilC18(250 mm×4.6 mm,5μm),流动相:乙腈-0.05%磷酸(15:85),检测波长:203 nm,流速为1 mL/min。结果:人参皂苷Re的回归方程为:Y=91273.9x+313 618г=0.999 8,(n=6)。线性范围为:52.46-314.76 mg/L,平均回收率为99.54%,RSD为0.38%(n=5)。结论:该方法具有简便、快速、准确,重现性好,与标准法比较无显著性差别,适合于参麦注射液含量的测定。     

西罗莫司纳米脂质载体固化制剂的制备

采用冷冻干燥法以微晶纤维素Avicel PH-101与聚乙烯吡咯烷酮(PVP)K30的混合物(4:1,w,%)为吸附性粉末固化西罗莫司纳米脂质载体分散液,并以再分散时间、平均粒径及分布、流动性和泄漏率等为指标,采用单因素试验优化固化处方。结果表明,3批按优化方法制备的西罗莫司纳米脂质载体固化制剂的休止角为(42.65±0.80)。,振实密度为(0.79±0.03)g/m1,且含量均匀度良好,再分散时间为(10.0±0.4)min,再分散液粒径(132.7+2.6)nm,分布系数0.297±0.01,ξ电位(-12.8±1.05)mV,冻干前后的泄漏率为(10.80±0.41)%。     

Improvement of the Prediction Power of the CoMFA and CoMSIA Models on Histamine H3 Antagonists by Different Variable Selection Methods

The aim of this study is to enhance the predictivity power of CoMFA and CoMSIA models by means of different variable selection algorithms. The genetic algorithm (GA), successive projection algorithm (SPA), stepwise multiple linear regression (SW-MLR), and the enhanced replacement method (ERM) were used and tested as variable selection algorithms. Then, the selected variables were used to generate a simple and predictive model by the multilinear regression algorithm. A set of 74 histamine H₃ antagonists were split into 40 compounds as a training set, and 17 compounds as a test set, by the Kennard-Stone algorithm. Before splitting the data, 17 compounds were randomly selected from the pool of the whole data set as an evaluation set without any supervision, pretreatment, or visual inspection. Among applied variable selection algorithms, ERM had noticeable improvement on the statistical parameters. The r² values of training, test, and evaluation sets for the ERM-MLR model using CoMFA fields were 0.9560, 0.8630, and 0.8460 and using the CoMSIA fields were 0.9800, 0.8521, and 0.9080, respectively. In this study, the principles of organization for economic cooperation and development (OECD) for regulatory acceptability of QSARs are considered.     

大鼠膝关节软骨不同染色方法的差异

背景：国内外学者对关节软骨对软骨进行了大量研究,需要用不同的染色方法对研究结果进行分析,但将多种染色方法同时应用于软骨研究及探讨染色机制的较少。 目的：探讨不同染色方法对大鼠膝关节软骨染色的优缺点。 方法：取正常大鼠膝关节软骨,行苏木精-伊红、番红O、阿尔辛蓝、甲苯胺蓝、番红-阿尔辛蓝、番红-固绿染色,观察软骨结构。 结果与结论：苏木精-伊红、番红O、甲苯胺蓝染色均可观察到潮线,分别为蓝色、红色和蓝色,番红O和甲苯胺蓝染色强度朝潮线方向增强,番红O染色对潮线的观察优于其他染色方法。苏木精-伊红染色关节软骨4层结构清晰,软骨细胞呈柱状排列,基质呈均匀呈嗜碱性染色。番红O染色显示4层结构层次清楚,基质深层染色最红。阿尔辛蓝染色显示,pH 1.0时软骨细胞周边部分被阿尔辛蓝强烈染色,pH 2.5时阿尔辛蓝染色较深。甲苯胺蓝染色显示组织结构层欠清,胞核染色清晰,胞浆几乎不着色,基质呈淡蓝紫色。番红-阿尔辛蓝染色显示软骨表面及基质呈不均一的红色,深层颜色较深,软骨细胞周围蓝染;番红-固绿染色显示软骨基质呈均匀的红色,软骨下骨呈绿色,软骨组织与骨组织分对比鲜明。提示上述各种方法均可观察到软骨的四层结构,但以番红O染色显示软骨各层和潮线结构最佳;苏木精-伊红染色观察软骨细胞形态变化较其他方法清楚。     

Facts and Fallacies About Measuring Blood Pressure in Rats

Although blood pressure can easily be measured in anesthetized rats by simply connecting a catheter to a pressure transducer, repeated measurements taken over long periods of time in awake rats are much more difficult to make. For chronic experiments two methods are now commonly used: direct recording from chronically-implanted arterial catheters, or indirect measurement with the tail-cuff method. Direct recording of intraarterial pressure can be done continuously and is more accurate, but technically more demanding. On the other hand, although tail-cuff measurements are less accurate, they do not require surgery and can be repeated almost indefinitely. With most tail-cuff methods the rats are preheated to dilate the tail vessels and thereby facilitate pulse detection, but with the new IITC photoelectric sensor indirect measurements of systolic as well as of mean arterial pressure can be made without external preheating. Even with a properly validated tail-cuff method, however, errors can still occur particularly when it is used to quantify modest blood pressure changes like those during development of hypertension, or following administration of vasoactive drugs. To safeguard against such errors, each laboratory should always validate its own tail-cuff method under uniform experimental conditions similar to those existing when the method is actually used. Additionally, all blood pressure differences thereby detected should be verified by direct measurement of intraarterial pressure in the same rats.     

A test of concordance between patient and psychiatrist valuations of multiple treatment goals for schizophrenia

Background While much discussion has been placed on the problem of poor compliance in the treatment of schizophrenia, there has been little discussion on the concordance between patients and psychiatrists, an important contributing factor to patient‐centred care. Objective To estimate the concordance between patients’ and psychiatrists’ (ordinal and cardinal) valuations of multiple goals for schizophrenia treatment and to illustrate the utility of the self‐explicated method in valuing a large number of treatment goals. Design Twenty treatment goals were identified during focus groups and literature review and were presented to patients and psychiatrists during structured interviews. Respondents were asked to rank the multiple treatment goals and rate them on a 5‐point Likert scale. Three scores were calculated based on the ranking (1–20), rating (Likert scale) (1–5) and a self‐explicated method estimated as the product of rating and ranking score (1–100). Concordance was tested using Spearman’s rho for overall ordinal rankings and via anova and F‐test for the cardinal values assigned to a specific treatment goal. Participants A total of 105 outpatients diagnosed with schizophrenia and 160 psychiatrists in Germany. Results Patient and psychiatrist values were concordant when the ordinal properties of their valuations were assessed by rating (ρ = 0.63; P = 0.002), ranking (ρ = 0.51; P = 0.02) and self‐explicated methods (ρ = 0.54; P = 0.01). Significant discordances were found when comparing the cardinal value placed on any given treatment goal using all three approaches, but the self‐explicated method produced a more discerning statistic. Relative to patients, psychiatrists significantly (P < 0.05) overvalued reduced lack of emotion, improved sexual pleasure and improved communication while undervaluing reuptake of activities of daily living, improved satisfaction and recovered capacity for work. Conclusions While there is an overall concordance between patients’ and psychiatrists’ valuation, significantly different valuations on specific goals can be identified. Here, psychiatrists tend to focus on ‘textbook’ outcomes, while patients are more concerned with functioning and living a normal life. This study also demonstrates the importance of comparing the concordance in treatment goals and the importance of preference‐based methods, such as the self‐explicated method, in the study of concordance.     

SERUM-FREE CULTURE OF RHESUS MONKEY EMBRYONIC STEM CELLS

Previous studies have shown that the maintenance and proliferation of undifferentiated rhesus monkey embryonic stem (rES) cells requires medium supplemented with fetal bovine serum (FBS). Due to the uncharacterized composition and variation in serum nature, the present study aimed to replace the serum-containing medium with a serum-free medium in the rES cell culture. The results showed that after the initial 48-h culture in the routinely used serum-containing medium, rES cells can grow and proliferate for a prolonged period in the serum-free medium composed of DMEM supplemented with a cocktail of BSA, IGF-1, TGF- &#102, bFGF, aFGF, estradiol, and progesterone. rES cells cultured in the serum-free medium maintained high level of alkaline phosphatase activity and OCT4 level. There was no indication of differentiation as judged by the marker gene expression of all three embryonic germ layers and trophoblast. In addition, serum-free culture would not affect the passage capacity and differentiation potential of rES cells. This work will facilitate the future study of induced differentiation of rES cells and other applications.