地塞米松与甲硝唑在防治根管治疗期间急症中的联合应用

目的:观察联合应用地塞米松与甲硝唑行根管治疗期间急症处理的效果。方法:对66例根管治疗期间急症发作的病例分别采用联合应用地塞米松与甲硝唑、单独使用地塞米松和单独使用甲醛甲酚的方法治疗,观察各组的差异。结果:地塞米松与甲硝唑联合应用组的治疗效果优于其他组,差异显著。结论:联合应用地塞米松与甲硝唑可有效治疗根管治疗期间急症。     

Identification of Endothelial Proteins by MALDI-MS Using a Compact Disc Microfluidic System

Vascular endothelial proteins have been analyzed using two-dimensional (2D) gel electrophoresis and subsequent mass spectrometry, with separate methods for the intervening sample preparations. Compact disc (CD) technology was found to be rapid, giving high overall yield both with ordinary Coomassie staining and with Sypro Ruby staining. Combined with automatic in-gel digestion, the CD technology has great capacity for large numbers of protein analysis, although for limited sample numbers, manual methods can give similar sequence coverage. In a test set of 48 samples, 45 proteins were identified using the CD preparation technique, 32 identified with higher sequence coverage using the CD technique, 7 with higher using ZipTips in a robotic workstation, and 5 with higher coverage using dried droplets of unpurified samples. In the process of these methodological comparisons, basic patterns for 116 endothelial proteins were defined, representing 297 separate protein spots on the 2D gels.     

Measurements of DNA and RNA in mammary gland homogenates by the ethidium bromide technique.

A rapid method of determining simultaneously DNA and RNA in mammary gland homogenates using the ethidium bromide technique is discussed. The method utilizes a quantitative extraction of DNA and RNA with 2.0m sodium chloride, SDS, and EDTA at pH 8.0. Assays of mammary gland RNA and DNA using previously published methods were compared with determinations using the ethidium bromide technique. While the fluorescence method gave lower values for RNA when compared to those obtained using the orcinol or absorbancy ratio (OD 260nm/280nm), DNA measurements agreed well with the values determined by the diphenylamine technique. Extinction coefficient data for total mammary gland RNA isolated using a modified phenol extraction procedure are also presented.     

Pharmacokinetic interaction with digoxin and glucocorticoids in rats detected by radio-immunoassay using a novel specific antiserum

We previously prepared a more specific antiserum (Antiserum-I) to digoxin (Dx) compared with commercially available anti-Dx antiserum (Antiserum-II), clinically used in the therapeutic drug monitoring of Dx. The aims of this study are to compare Dx disposition kinetics by radio-immunoassay (RIA) using Antiserum-I and Antiserum-II, and evaluate the drug-drug interaction with Dx and glucocorticoids in rats. When Dx metabolites were added to rat serum containing Dx, the recovery ratios using Antiserum-I showed 100 to 110% and were remarkably lower than those using Antiserum-II. In rats, serum concentration-time courses of Dx after a single i.v. or p.o. administration of Dx (0.017 mg/kg) by RIA using Antiserum-I were much lower than those using Antiserum-II. The area under the concentration-time course of Dx was significantly lower than that using Antiserum-II and the total body clearance values were significantly higher, while an obvious change of bioavailability was not observed. When using Antiserum-I, rats twice and six times pretreated with dexamethasone (75 mg/kg/day, i.p.) and prednisolone (69 mg/kg/day, i.p.), respectively, showed significant change of the pharmacokinetic parameters of Dx compared with the control rats. In contrast, using Antiserum-II, it took three and nine times of pretreatment with dexamethasone and prednisolone, respectively, to significantly change the parameters of Dx. In conclusion, these results demonstrate that Antiserum-I is very useful not only to more precisely monitor serum Dx levels, but also to determine earlier the drug-drug interaction with glucocorticoids than Antiserum-II.     

Comparison between agarose gel electrophoresis and capillary electrophoresis for variable numbers of tandem repeat typing of Mycobacterium tuberculosis

Variable numbers of tandem repeat (VNTR) typing of Mycobacterium tuberculosis was performed on 54 strains including 23 strains derived from 9 outbreaks. PCR amplicon sizes of 12 mycobacterial interspersed repetitive unit tandem repeat loci were measured using both agarose gel electrophoresis and capillary electrophoresis. Similarities using agarose gel electrophoresis of Euclidian distances among the 23 strains derived from the 9 outbreaks were significantly lower than that using capillary electrophoresis (Wilcoxon signed ranks test, P < 0.01). By clustering analysis using unweighted pair group method using arithmetic averages, all of the 23 strains derived from the 9 outbreaks were each clustered with more than 90% similarities based on the distance using capillary electrophoresis. In contrast, differential clusters with more than 90% similarity were observed with only 7 strains derived from 3 outbreaks when analyzed by agarose gel electrophoresis. These results indicated that measurement of PCR amplicon size of tandem repeat loci should be carried out using capillary electrophoresis and that agarose gel electrophoresis is not suitable for clustering analysis of M. tuberculosis VNTR typing.     

Direct detection of herceptin/trastuzumab binding on breast tissue sections.

The protooncogene product HER-2/neu is the target of the humanized monoclonal antibody trastuzumab (Herceptin). Several tests are used clinically to identify patients with HER-2/neu overexpression based on evaluation by pathologists of gene amplification by fluorescence in situ hybridization or protein expression using immunohistochemistry (IHC). A simple technique has been developed for staining formalin-fixed, paraffin-embedded breast cancer tissue using unmodified Herceptin/trastuzumab as the primary antibody. Results were compared with staining with the commercial kit, HercepTest, as well as with polyclonal anti-HER-2/neu antibodies and with biotinylated trastuzumab. These procedures were tested using four breast cancer microarrays. There were 854 cores that were stained with all four antibodies, representing 325 cases. A standard 4-point scoring system (0-3) was used. A total of 156 cases (48%) were scored as 0 by all the methods used and 31 (9.5%) were positive (3+) by all methods. Of interest, three cases scored negative using polyclonal anti-HER-2/neu antibodies but were positive using unmodified trastuzumab. To clarify this discrepancy, whole sections of tumors were examined with both antibodies using double labeling. There were some tumors that demonstrated a mosaic pattern of staining with neighboring cells or groups of cells stained exclusively with one antibody or the other. These results demonstrate that unmodified humanized or human therapeutic antibodies could be used for preclinical testing or in a clinical laboratory setting for IHC-based selection of patients for treatment, and results of such selection could be different from those obtained using polyclonal antibody-based IHC procedure.     

Transfection of embryonal carcinoma cells at high efficiency using liposome-mediated transfection

Embryonal carcinoma (EC) cells are recognized as an excellent model system for studying the early stages of mammalian development. Many studies performed with EC cells involve transient transfection with promoter/reporter gene constructs and/or mammalian expression vectors. One of the limitations of working with EC cells is their inability to be transfected at high efficiency. In most cases, EC cells are transfected using the calcium phosphate method. The objective of this study was to identify protocols and culture conditions that significantly increase the transfection efficiency of EC cells. F9 EC cells were used for this purpose, because they are the EC cell line studied most commonly. We show that the transfection efficiency of F9 EC cells using the calcium phosphate method is less than 5%; whereas, their transfection efficiency can be improved approximately 15-fold using optimized culture conditions and liposome-based transfection reagents. Specifically, we demonstrate that more than 50% of F9 EC cells can be transfected using LipofectAMINE 2000. In addition to higher levels of transfection, there is much less plate-to-plate variation with liposome-based reagents as compared to transfection with calcium phosphate. Interestingly, transfection efficiency using these reagents was found to be inversely related to cell density. This contrasts sharply with the recommendation that transfection with LipofectAMINE 2000 or LipofectAMINE in conjunction with the PLUS reagent be performed at high cell densities. Given the improvements in transfection efficiency reported here, it will now be possible to perform studies with F9 EC cells that require transfection at significantly higher levels than that achieved using the calcium phosphate method. Overall, the highest transfection efficiencies were consistently obtained using LipofectAMINE 2000.     

Systematic Position of the Rhoipteleaceae: Evidence from Nucleotide Sequences of rbc L Gene

Systematic position of the Rhoipteleaceae was investigated using nucleotide sequences of the chloroplast gene rbcL. Two sets of parsimony analyses were conducted, one using Bauera as the outgroup and the other using Cercidiphyllum-Liquidambar as outgroups. The results of these analyses are consistent. Rhoiptelea is allied with sampled Jug-landaceous genera in the rbcL phylogeny, suggesting its close relationship with the Juglandaceae.     

Comparative analysis of crystallinity changes in cellulose I polymers using ATR-FTIR, X-ray diffraction, and carbohydrate-binding module probes

Cotton fiber cellulose is highly crystalline and oriented; when native cellulose (cellulose I) is treated with certain alkali concentrations, intermolecular hydrogen bonds are broken and Na-cellulose I is formed. At higher alkali concentrations Na-cellulose II forms, wherein intermolecular and intramolecular hydrogen bonds are broken, ultimately resulting in cellulose II polymers. Crystallinity changes in cotton fibers were observed and assigned using attenuated total reflectance Fourier transform infrared (ATR FT-IR) spectroscopy and X-ray diffraction (XRD) subsequent to sodium hydroxide treatment and compared with an in situ protein-binding methodology using cellulose-directed carbohydrate-binding modules (CBMs). Crystallinity changes observed using CBM probes for crystalline cellulose (CBM2a, CBM3a) and amorphous cellulose (CBM4-1, CBM17) displayed close agreement with changes in crystallinity observed with ATR-FTIR techniques, but it is notable that crystallinity changes observed with CBMs are observed at lower NaOH concentrations (2.0 mol dm(-3)), indicating these probes may be more sensitive in detecting crystallinity changes than those calculated using FTIR indices. It was observed that the concentration of NaOH at which crystallinity changes occur as analyzed using the CBM labeling techniques are also lower than those observed using X-ray diffraction techniques. Analysis of crystallinity changes in cellulose using CBMs offers a new and advantageous method of qualitative and quantitative assessment of changes to the structure of cellulose that occur with sodium hydroxide treatment.     

Image quality assessment with dose reduction using high kVp and additional filtration for abdominal digital radiography

PurposeDose reduction using additional filters with high kilovoltage peak (kVp) for abdominal digital radiography has received much attention recently. We evaluated image quality with dose reduction in abdominal digital radiography by using high kVp and additional copper filters at a tertiary hospital.MethodsBetween June 2016 and July 2016, 82 patients underwent abdominal digital radiography using 80 kVp in X-ray room 1 and 82 were imaged using 92 kVp with 0.1-mm copper filtration in X-ray room 2. The effective dose was calculated using a PC-based Monte Carlo program. Image quality of the abdominal radiography acquired in the two rooms was evaluated using a five-point ordinal scale, as well as the signal-to-noise and contrast-to-noise ratios.ResultsThe mean effective dose decreased by 25.8% and 25.7% for the supine and standing positions, respectively, when abdominal digital radiography using 92 kVp with 0.1-mm copper filtration was performed. In the 20 patients who performed abdominal digital radiography twice in each room, visual grading scores for visualisation of psoas outlines and kidney outlines are higher in room 1. However, there was no statistical significant difference of visual grading scores among the 124 patients who underwent only one abdominal radiography in the room 1 or 2 (P > 0.05).ConclusionsDose reduction for abdominal digital radiography can be achieved with comparable image quality by performing abdominal digital radiography using 92 kVp with 0.1-mm copper filtration, despite the higher AEC dose.     

Fabrication of nanofibers with antimicrobial functionality used as filters: protection against bacterial contaminants

A comparative study of antimicrobial activity is done using three different electrospun nanofibers namely-CA, PAN, and PVC used as control and with various amounts of AgNO(3) being treated with UV-irradiation leading to the enhancement of silver nanoparticles. DMF is used as the common solvent which helps to undergo spontaneous slow reduction at room temperature to form silver nanoparticles followed by UV-irradiation using a 400 W source. The time required for the formation of silver nanoparticles is short and they are more or less well dispersed with few such aggregates. The presence of silver nanoparticles is confirmed using various characterization techniques. The antimicrobial activity is studied using nanofibers with fabricated functionality.     

马尾树科的系统位置:来自rbcL基因核苷酸序列的证据

本文根据叶绿体基因rbcL的核苷酸序列证据讨论了单型科——马尾树科的系统关系。数据矩 阵中包括了马尾树科及其它“高等”金缕梅类各科,并分别利用Cunoniaceae科的bauera属,“低等”金缕梅类植物连香树属和枫香树属作为外类群进行分支分析。两种分析的结果是一致的。在rbcL基因树上,马尾树属和被用于分析的胡桃科各属紧密地结合在一起,支持马尾树科和胡桃科有非常近的亲缘关系。     

Quantitation of phosphotyrosine signals in human prostate cell adhesion sites

A simple method is described for the quantitation of phosphotyrosine signaling in human prostate cell cultures. The phosphotyrosine signals are observed by standard immunohistochemistry techniques, and the resulting digital images are analyzed using the Scion image software program. The signals within the cell adhesion sites are quantitated using the density slice and particle analysis features of the software. The immunohistochemistry results are compared with detection of phosphotyrosine signals using a standard Western blotting procedure with whole cell lysates. The resulting data is converted into graphs using the Sigma Plot Program. This method is illustrated using damage-induced signaling within cell adhesion sites after a low dose of ionizing radiation.     

Total Counts Versus Line Transects for Estimating Abundance of Small Gopher Tortoise Populations

Abstract: Status and trends of gopher tortoise (Gopherus polyphemus) populations are a critical information need for natural resource managers, researchers, and policy makers. Many tortoise populations are small and isolated, which can present challenges for deriving population estimates. Our objective was to compare abundance and density estimates for a small tortoise population derived using a total burrow count versus estimates obtained with line transect distance sampling (LTDS) using repeated surveys. We also compared results of the 2 survey methods using standard burrow-to-tortoise correction factors versus assessing occupancy of all burrows with a camera scope. In addition, we compared LTDS data obtained using a compass and measuring tape to define transects to those obtained using a Global Positioning System (GPS) and Personal Data Assistant (PDA) field computer to navigate transects. Line transect distance sampling with repeated surveys (both with a measuring tape and compass and with a GPS—PDA) yielded sufficient observations of tortoises to calculate population estimates. From 18% to 31% of burrows were occupied by tortoises as determined with the burrow camera. We found 25 burrows during the LTDS survey that we did not find in the total count survey, which demonstrated that the assumption of 100% detection for the total count was not met; hence, density or abundance measurements derived with this method were underestimates. We recommend using GPS—PDA technology, scoping all burrows detected, and using LTDS with repeated surveys to estimate abundance and density for small gopher tortoise populations.     

Digoxigenin-labeled probe for rapid identification of nisinogenic Lactococcus lactis strains

From the nisZ gene sequence, a non-radioactive digoxigenin-labeled DNA probe, was tested for detection of nisin-producing strains using polymerase chain reaction amplification. The digoxigenin-labeled DNA probe clearly discriminated between nisin-producing and non-producing strains with a high degree of sensitivity and specificity. By agarose gel electrophoresis, 1.4 ng of nisin DNA was detected using the digoxigenin-labeled DNA probe compared with 11 ng using direct polymerase chain reaction amplification. A colony hybridization method using digoxigenin-labeled DNA to selectively detect nisinogenic bacteria showed that the nis-probe was specific and did not react with any other non-bacteriocinogenic and non-nisinogenic strains.     

Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers

A method for apolipoprotein (apo) E genotyping was developed using the polymerase chain reaction (PCR) with allele-specific oligonucleotide primers (ASP). Synthetic oligonucleotides with base-pair mismatches at the 3' terminus were used as primers to amplify the apoE gene in subjects previously phenotyped using isoelectric focusing (IEF). Complementary primer-allele combinations were specifically amplified by PCR, together with a control pair of primers specific to the human prothrombin gene. Identification of genotype by PCR using ASP was consistent with the phenotypes that were determined by IEF for 14 healthy normolipidemic subjects. These results were achieved using DNA isolated from buccal epithelial cells obtained from a mouthwash or DNA extracted from leukocytes. Genotype identification required analysis of the PCR products on an ethidium-stained agarose gel, yielding results 3 h after DNA extraction. In comparison with other current methods, PCR using ASP is suggested as a rapid and simple noninvasive technique for determining population apoE allelic distribution.     

Comparison of the potentiometric, (31)P NMR, and zero-point titration methods of determining ionized magnesium in erythrocytes

A simple and rapid method of determining ionized magnesium in erythrocytes using a potentiometric clinical analyzer, Microlyte 6 (Kone, Finland), was investigated. The erythrocyte cell membranes were destroyed using ultrasound. The results were compared with those obtained with the (31)P nuclear magnetic resonance spectroscopy method and the zero-point titration method using atomic absorption spectrometry. The results obtained from potentiometry and from the other methods did not differ significantly (Student t test, alpha = 0.01). Total magnesium concentration was determined using atomic absorption spectrometry.     

650例疑似细菌性阴道病BV患者阴道分泌物检测分析

目的探讨生化检测法在细菌性阴道病（BV）诊断中的应用价值。方法对绍兴市人民医院妇科门诊650例阴道分泌物进行常规及生化检测。结果在650例疑似细菌性阴道病患者中,以“唾液酸苷酶阳性＋过氧化氢阳性”为诊断标准,共检出BV患者483例（占总数的74．31％）;使用Amsel法检出细菌性阴道病患者481例（占总数的74％）。使用生化检测法和Amsel法检出细菌性阴道病的阳性率差异无统计学意义（x2=0．53,P〉O．05）,二者的符合率为94．15％（612／650）,具有很高的一致性。结论生化检测准确、快速,具有较高的临床应用价值。     

Purification of coagulation factor VIII using chromatographic methods. Direct chromatography of plasma in anion exchange resins

Coagulation factor VIII (FVIII) concentrates are used in the treatment of patients with Hemophilia A. Human FVIII was purified directly from plasma using anion exchange chromatography followed by gel filtration. Three Q-Sepharose resins were tested, resulting in 40% recovery of FVIII activity using Q-Sepharose XL resin, about 80% using Q-Sepharose Fast Flow and 70% using the Q-Sepharose Big Beads. The vitamin K-dependent coagulation factors co-eluted with FVIII from the anion exchange columns. In the second step of purification, when Sepharose 6FF was used, 70% of FVIII activity was recovered free from vitamin K-dependent factors.