四位数码翻号床牌在复苏室手术患者床位标识中的应用体会

目的：探讨四位数码翻号床牌在复苏室手术患者床位标识的方法。方法巧用四位数码翻号床牌对我院所有需进入复苏室的手术患者进行床位标识。结果在使用过程中，翻号床牌操作简单方便，标识清楚，可准确和直观的看到复苏病人的床位标识，分辨率极高。结论四位数码翻号床牌，能准确标识手术复苏患者的床位号，便于医护人员三查七对，避免差错事故的发生，提高了复苏室抢救效率。     

Stroop interference associated with efficient reading fluency and prelexical orthographic processing

The Stroop Color–Word Test involves a dynamic interplay between reading and executive functioning that elicits intuitions of word reading automaticity. One such intuition is that strong reading skills (i.e., more automatized word reading) play a disruptive role within the test, contributing to Stroop interference. However, evidence has accumulated that challenges this intuition. The present study examined associations among Stroop interference, reading skills (i.e., isolated word identification, grapheme-to-phoneme mapping, phonemic awareness, reading fluency) measured on standardized tests, and orthographic skills measured on experimental computerized tasks. Among university students (N = 152), correlational analyses showed greater Stroop interference to be associated with (a) relatively low scores on all standardized reading tests, and (b) longer response latencies on orthographic tasks. Hierarchical regression demonstrated that reading fluency and prelexical orthographic processing predicted unique and significant variance in Stroop interference beyond baseline rapid naming. Results suggest that strong reading skills, including orthographic processing, play a supportive role in resolving Stroop interference.     

Direct Identification of Enteroviruses in Cerebrospinal Fluid of Patients with Suspected Meningitis by Nested PCR Amplification

Enteroviruses, the most common human viral pathogens worldwide, have been associated with serous meningitis, encephalitis, syndrome of acute flaccid paralysis, myocarditis and the onset of diabetes type 1. In the future, the rapid identification of the etiological agent would allow to adjust the therapy promptly and thereby improve the course of the disease and prognosis. We developed RT-nested PCR amplification of the genomic region coding viral structural protein VP1 for direct identification of enteroviruses in clinical specimens and compared it with the existing analogs. One-hundred-fifty-nine cerebrospinal fluids (CSF) from patients with suspected meningitis were studied. The amplification of VP1 genomic region using the new method was achieved for 86 (54.1%) patients compared with 75 (47.2%), 53 (33.3%) and 31 (19.5%) achieved with previously published methods. We identified 11 serotypes of the Enterovirus species B in 2012, including relatively rare echovirus 14 (E-14), E-15 and E-32, and eight serotypes of species B and 5 enteroviruses A71 (EV-A71) in 2013. The developed method can be useful for direct identification of enteroviruses in clinical material with the low virus loads such as CSF.     

鲍曼不动杆菌耐药性快速检测方法的建立与评估

目的评估对实时荧光定量聚合酶链反应(PCR)快速检测鲍曼不动杆菌耐药性方法的效果。方法设计鲍曼不动杆菌(ATCC 19606)外膜蛋白A特异性探针,采用实时荧光定量PCR准确测定63株鲍曼不动杆菌经一定浓度庆大霉素、米诺环素、美罗培南、左氧氟沙星和头孢哌酮/舒巴坦孵育后的生长情况,比较抗菌药物孵育0和6h后鲍曼不动杆菌基因组DNA量,判断鲍曼不动杆菌耐药性,并与微量肉汤稀释法结果进行对比。结果建立的实时荧光定量PCR检测鲍曼不动杆菌耐药性方法与微量肉汤稀释法一致性高,Kappa值为0.879,P0.05。以微量肉汤稀释法为参考方法,实时荧光定量PCR的符合率达93.97%,灵敏度为92.53%,特异性为95.74%,阳性预测值为96.41%,阴性预测值为91.22%,Youden指数为0.88。结论实时荧光定量PCR与微量肉汤稀释法结果有高度一致性,且前者灵敏度高、特异性强、检测速度快,可用于快速检测鲍曼不动杆菌的耐药性。     

从外观性状识别酸枣仁伪品

探讨酸枣仁及其常见伪品的性状鉴别,确保临床用药的安全、有效。取酸枣仁及其常见伪品,对外观性状进行比对讨论发现酸枣仁及其伪品在外观性状有明显区别。性状鉴别方法能有效地识别酸枣仁及其伪品。     

Engineering of a recombinant trivalent single-chain variable fragment antibody directed against rabies virus glycoprotein G with improved neutralizing potency

Human and equine rabies immunoglobulins are currently available for passive immunization against rabies. However, these are hampered by the limited supply and some drawbacks. Advances in antibody engineering have led to overcome issues of clinical applications and to improve the protective efficacy. In the present study, we report the generation of a trivalent single-chain Fv (scFv50AD1-Fd), that recognizes the rabies virus glycoprotein, genetically fused to the trimerization domain of the bacteriophage T4 fibritin, termed ‘foldon’ (Fd). scFv50AD1-Fd was expressed as soluble recombinant protein in bacterial periplasmic space and purified through affinity chromatography. The molecular integrity and stability were analyzed by polyacrylamide gradient-gel electrophoresis, size-exclusion chromatography and incubation in human sera. The antigen-binding properties of the trimeric scFv were analyzed by direct and competitive-ELISA. Its apparent affinity constant was estimated at 1.4 ± 0.25 × 10⁹ M⁻¹ and was 75-fold higher than its monovalent scFv (1.9 ± 0.68 × 10⁷ M⁻¹). The scFv50AD1-Fd neutralized rabies virus in a standard in vitro and in vivo neutralization assay. We showed a high neutralization activity up to 75-fold compared with monovalent format and the WHO standard serum. The gain in avidity resulting from multivalency along with an improved biological activity makes the trivalent scFv50AD1-Fd construct an important reagent for rabies protection. The antibody engineering approach presented here may serve as a strategy for designing a new generation of anti-rabies for passive immunotherapy.     

Formulation and optimization of potassium iodide tablets

The use of potassium iodide (KI) as a protective agent against accidental radioactive exposure is well established. In this study, we aimed to prepare a KI tablet formulation using a direct compression method. We utilized Design of Experiment (DoE)/mixture design to define the best formulation with predetermined physical qualities as to its dissolution, hardness, assay, disintegration, and angle of repose. Based on the results from the DoE, the formulation had the following components (%w/w): Avicel 48.70%, silicon dioxide 0.27%, stearic acid (1.00%), magnesium stearate 2.45%, and dicalcium phosphate 18.69%, in addition to potassium iodide 28.89% (130 mg/tablet). This formulation was scaled-up using two tablet presses, a single-punch press and a rotary mini tablet press. The final scaled-up formulation was subjected to a variety of quality control tests, including photo-stability testing. The results indicate that potassium iodide tablets prepared by a rotary mini tablet press had good pharmaceutical characteristics and a shelf-life of 25 days when stored at room temperature protected from light.     

From the Cover: Searching molecular structure databases with tandem mass spectra using CSI:FingerID

Metabolites provide a direct functional signature of cellular state. Untargeted metabolomics experiments usually rely on tandem MS to identify the thousands of compounds in a biological sample. Today, the vast majority of metabolites remain unknown. We present a method for searching molecular structure databases using tandem MS data of small molecules. Our method computes a fragmentation tree that best explains the fragmentation spectrum of an unknown molecule. We use the fragmentation tree to predict the molecular structure fingerprint of the unknown compound using machine learning. This fingerprint is then used to search a molecular structure database such as PubChem. Our method is shown to improve on the competing methods for computational metabolite identification by a considerable margin.Metabolites, small molecules that are involved in cellular reactions, can provide detailed information about cellular state. Untargeted metabolomic studies may use NMR or MS technologies, but liquid chromatography followed by MS (LC/MS) can detect the highest number of metabolites from minimal amounts of sample (1, 2). Untargeted metabolomics comprehensively compares the mass spectral intensities of metabolite signals (peaks) between two or more samples (3, 4). Advances in MS instrumentation allow us to simultaneously detect thousands of metabolites in a biological sample. Identification of these compounds relies on tandem MS (MS/MS) data, produced by fragmenting the compound and recording the masses of the fragments. Structural elucidation remains a challenging problem, in particular for compounds that cannot be found in any spectral library (1): In total, all available spectral MS/MS libraries of pure chemical standards cover fewer than 20,000 compounds (5). Growth of spectral libraries is limited by the unavailability of pure reference standards for many compounds.In contrast, molecular structure databases such as PubChem (6) and ChemSpider (7) contain millions of compounds, with PubChem alone having surpassed 50 million entries. Searching in molecular structure databases using MS/MS data is therefore considered a powerful tool for assisting an expert in the elucidation of a compound. This problem is considerably harder than the fundamental analysis step in the shotgun proteomics workflow, namely, searching peptide MS/MS data in a peptide sequence database (8): Unlike proteins and peptides, metabolites show a large structural variability and, consequently, also large variations in MS/MS fragmentation. Computational approaches for interpreting and predicting MS/MS data of small molecules date back to the 1960s (9): Due to the unavailability of molecular structure databases at that time, structure libraries were combinatorially generated and then “searched” using the experimental MS/MS data. “Modern” methods for this question have been developed since mid-2000. Particular progress has been made for restricted metabolite classes such as lipids (5), but as with peptides, results cannot be generalized to other metabolite classes. For the general case, several strategies have been proposed during recent years, including simulation of mass spectra from molecular structure (10, 11), combinatorial fragmentation (12–17), and prediction of molecular fingerprints (18, 19).Searching in a molecular structure database is clearly limited to those compounds present in the database. Fragmentation trees have been introduced as a means of analyzing MS/MS data without the need of any (structural or spectral) database (20–22). In this paper, the term “fragmentation tree” is exclusively used to refer to the graph-theoretical concept introduced in ref. 20, not “spectral trees” that describe the dependencies of multiple MS measurements; see Vaniya and Fiehn (23) for a review. In more detail, our fragmentation trees are predicted from MS/MS data by an automated computational method such that peaks in the MS/MS spectrum are annotated with molecular formulas of the corresponding fragments, and fragments are connected via assumed losses. Clearly, there exist other approaches with the broad aim of identifying metabolites, such as network-based methods (24–26) and combined approaches (27); see Hufsky et al. (28) for a review of computational methods in MS-based metabolite identification.It is undisputed that MS/MS data alone are insufficient for full structural elucidation of metabolites. We argue that elucidation of stereochemistry is currently beyond the power of automated search engines, so we try to recover the correct constitution (bond structure) of the query molecule, that is, the identity and connectivity (with bond multiplicities) of the atoms, but no stereochemistry information. Throughout this paper, we refer to the constitution of the molecule as its structure. In practice, orthogonal information is usually available, both analytical (retention time, ion mobility drift time, infrared and UV spectroscopy, and NMR data) and on the experimental setup (extraction procedure and organism) (29, 30). We assume that this information is not presented to the search engines but rather used in a postprocessing step to manually select the best solution from the output list of the engine. This is comparable to the everyday use of search engines for the internet.Here, we present CSI (Compound Structure Identification):FingerID for searching a molecular structure database using MS/MS data. Our method combines computation and comparison of fragmentation trees with machine learning techniques for the prediction of molecular properties of the unknown compound (19). Our method shows significantly increased identification rates compared with all existing state-of-the-art methods for the problem. CSI:FingerID is available at www.csi-fingerid.org/. Our method can expedite the identification of metabolites in an untargeted workflow for the numerous cases where no reference measurements are available in spectral libraries.     

成人表皮干细胞和汗腺细胞的体外分离和培养

背景：人表皮干细胞和汗腺细胞的分离培养及鉴定,为探讨人表皮干细胞再生汗腺的可行性打下基础。目的：探讨体外分离培养人表皮干细胞及汗腺细胞的有效方法。方法：采集不同年龄段的泌尿外科患者术后包皮组织,充分清洗消毒后去除皮下组织,将其修剪成0.5cm×0.5cm的皮片,用Ⅳ型胶原纯化、富集成人表皮干细胞,倒置相差显微镜观察细胞形态：使用免疫荧光染色对成人表皮干细胞表型进行分析;CCK-8检测细胞的增殖曲线;采用胶原酶消化法从人全层无损伤皮肤中分离提取汗腺细胞,并进行扩增和鉴定。结果：倒置相差显微镜下见分离培养的成人表皮干细胞呈卵圆形、细胞之间紧密相连,呈铺路石状,免疫荧光染色显示细胞表达CK19、β1整合素。倒置相差显微镜下见汗腺细胞呈扁平多角形,表达汗腺细胞标志细胞CK7、18、19及CEA。结论：本实验结果说明胶原酶消化法分离培养成人表皮干细胞及汗腺样细胞是可行的。