Clinical role and importance of fluorescence in situ hybridization method in diagnosis of H pylori infection and determination of clarithromycin resistance in H pylori eradication therapy

H pylori is etiologically associated with gastritis, gas-tric and duodenal ulcers, gastric adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma. Eradicating H pylori may convert rapidly the outcome of related diseases with the use of more accurate diagnostic molecular tests. Indeed some of the tests cannot give the evidence of current infection; H pylori can be detected by noninvasive and invasive methods, the latter requiring an endoscopy. Eradication failure is a big problem in H pylori infection. Recently, clarithromycin resistance in H pylori strains is increasing and eradicati-on therapy of this bacterium is becoming more difficult. Molecular methods have frequently been applied besides phenotypic methods for susceptibility testing to detect clarithromycin resistance due to mutations in the 2143 and 2144 positions of 23S rRNA gene. Fluorescence in situ hybridization (FISH) method on paraffin embedded tissue is a rapid, accurate and cost-effective method for the detection of H pylori infection and to determine clarithromycin resistance within three hours according to the gold standards as a non-culture method. This method can also be applied to fresh biopsy samples and the isolated colonies from a culture of H pylori, detecting both the culturable bacillary forms and the coccoid forms of H pylori, besides the paraffin embedded tissue secti-ons. This technique is helpful for determining the bac-terial density and the results of treatment where clarith-romycin has been widely used in populations to increase the efficacy of the treatment and to clarify the treatment failure in vitro.

Adaptive hp finite element method for fluorescence molecular tomography with simplified spherical harmonics approximation

Recently,the simplified spherical harmonics equations(SP)model has at tracted much att entionin modeling the light propagation in small tissue ggeometriesat visible and near-infrared wave-leng ths.In this paper,we report an eficient numerical method for fluorescence moleeular tom-ography(FMT)that combines the advantage of SP model and adaptive hp finite elementmethod(hp-FEM).For purposes of comparison,hp-FEM and h-FEM are,respectively applied tothe reconstruction pro cess with diffusion approximation and SPs model.Simulation experiments on a 3D digital mouse atlas and physical experiments on a phantom are designed to evaluate thereconstruction methods in terms of the location and the reconstructed fluorescent yield.Theexperimental results demonstrate that hp-FEM with SPy model,yield more accurate results thanh-FEM with difusion approximation model does.The phantom experiments show the potentialand feasibility of the proposed approach in FMT applications.

A TOTAL CONTENT X—RAY FLUORESCENCE METHOD FOR COPPER PROSPECTING

A new method is proposed to prospect copper deposits with portable XRF analyzer. The method is based on the close relation between Cu and the chalcophile elements or some other elements in the geochemical anomalies of a Cu deposit. Applications of the technique in Northeast China are presented.

Influence enhancement effect of bi-frequency ultrasonic irradiation by TA fluorescence method

Based on a previous research of cavitation effect under bi-frequency ultrasound irradiation, this paper studies bi-frequency irradiations with similar experimental settings. The additional irradiation sources with frequencies of 1.04MHz, 0.8MHz and 1.7MHz are individually combined with the main ultrasonic irradiation source with frequency of 28kHz to form bi-frequency ultrasonic irradiation. The intensity of 28kHz irradiation was fixed at 12.5W/cm^2, while the intensity of the ultrasound at the other three frequencies is varied from1 W/cm^2 to 18 W/cm^2. It turns out that under the influence of the bi-frequency irradiation, the fluorescence intensity is obviously greater than the sum of those at individual frequencies. So the frequency of the additional sonication strikingly influences the fluorescence enhancement effect. For example, the fluorescence enhancement effect of 1.04MHz is stronger than that of 1.7MHz, and the enhancement effect of 0.8MHz is further stronger than that of 1.04MHz. Under the sonic intensity of (7.9)W/cm^2, the fluorescence intensity of 1.04MHz is approximately twice that of 1.7MHz while the fluorescence intensity of 0.8MHz is approximately 1.5 times that of 1.04MHz.

Microstructure and UV Fluorescence of Amorphous I/Au Composite Nanofilms Based on LVDCS Method

The amorphous I/Au composite nanofilms were prepared by low vacuum direct current sputtering(LVDCS) method. The optimized preparation technologies contain growth pressure, time, gaseous environment and annealing conditions. The maximum fluorescence emission(λemmax) of I/Au nanofilms was observed at wavelength of 375 nm, and the intensity of fluorescence emission peak of annealed I/Au films was smaller than that of unannealed one due to fewer amorphous Au nanoparticles, caused by annealing treatment. In the UV-Vis absorption spectra, the intensity of UV-Vis absorption peak of annealed I/Au nanofilms is larger than that of the unannealed one. This work also developed a new way to grow I/Au composite fluorescent thin films.

Study on State of Terbium Ion in Liposome by One order Derivative Fluorescence Quenching Method

The state of Tb3+ is investigated in liposome. When the concentration of PC is below CMC (critical micell concentration), most of Tb3+ is associated with PC, the binding constant is about 3.35×103 L/mol. When the concentration of PC is beyond CMC, most of Tb3+ is dimerized, the dimerization constant is about 3.92×104L/mol. In PC?CH?H2O system, the binding constant of Tb3+?CH complex 2.93×104L/mol is obtained.

A New Fluorescence Method for the Measurement of Intracellular Free Rare Earth Ions： Fura－2 as an Indicator

Fura-2 was first used to monitor intracellular RE(3+). The characterizations of RE3+-Fura-2 interactions in solutions simulating intracellular ionic composition show that La3+ and Y3+ both form 1:1 RE(3+)-fura-2 complexes. The apparent dissociation constants are 1.61×10(-7)M,and 4.04×10(-7)M,respectively. The mouse myeloma(SP2/0) cells exposed to free RE(3+) or free RE(3+) and some small molecules in body were Studied by single cell canon measurement system.The results support the assumption that RE(3+) ions cannot enter the cytoplasm of living cells, and the small molecules used in our experiments couldn't improve the permeability of RE(3+)ions.

New HPLC Method with Experimental Design and Fluorescence Detection for Analytical Study of Antihypertensive Mixture,Amlodipine and Valsartan

New HPLC method was developed for determination of amlodipine and valsartan in their binary mixture as a part of routine control of combined formulations. The method was validated to meet official requirements including selectivity, stability, linearity, precision and accuracy. Chromatography was carried out using a LiChrospher RP-18 column, a mixture containing acetonitrile, phosphate buffer of pH 3.5 and methanol (45:45:10, v/v/v) and new fluorescence detection at 255 nm for excitation and 448 nm for emission. The effect of methanol content, pH of the buffer, flow rate, detection wavelengths and column temperature was estimated in robustness study, according to a plan defined by the Plackett-Burman design. For identification of significant effects, both graphical and statistical methods were used. Ro-bustness for dissolution test was checked estimating the effects of paddle speed, temperature and pH of dissolution medium. The method was proved to complying with all official guidelines. Therefore, it is suitable for determination of amlodipine and valsartan in their binary mixtures for different analytical and pharmaceutical purposes.

Determination of Bisoprolol Fumarate by Fluorescence Quenching Method

[Objectives]To establish a new method for indirect determination of bisoprolol fumarate based on fluorescence quenching technology.[Methods]In ammonia water and ammonium chloride buffer solution at pH=9.2,whenλexcitation=277 nm andλemission=596 nm,with the increase of CCu2+,the fluorescence signal intensity of bisoprolol fumarate weakened,and the difference between the fluorescence intensity of bisoprolol fumarate itself and the fluorescence intensity of the test solution after the quencher Cu2+was added(ΔF)and Cbisoprolol fumarate showed a good linear relationship.[Results]In the range of 15.39-76.93μg/mL,ΔF=146.7 Cbisoprolol fumarate+482.1,r=0.9988,and the detection limit is 0.1391μg/mL.[Conclusions]The fluorescence quenching method has been applied to the determination of actual samples with a recovery rate of 99.9%and an RSD of 2.7%.The results are satisfactory.

A FLUORESCENCE QUENCHING IMNUNOASSAY METHOD FOR DETERMINATION OF TRACE ALBUMIN USING UNLABELED EUROPIUM CHELATE

A fluoroimmunoassay method using unlabeled europium chelate is described.The principle is similar to that of fluoroimmunoassay method using lanthanide chelate as labels.The procedure is simple because labeling process is omitted.The detection limit is about 10^(10) mol/L antigen.The relative standard deviation of immunoassay is less than 10%.The recoveries of human serum albumin and estradiol protein conjugate are 96-105% and 111% respectively.

Determination of Anionic Surfactants in Environmental Water Samples Using Fluorescence Enhancement Method Based on Fe_3O_4@PAA-RB Fluorescent Nanoparticles

[ Objective] The study aims at developing a novel fluorescence enhancement method to determine anionic surfactants. [ Method] Based on Fe3O4 @ PAA-RB fluorescent nanoparticles as fluorescent probes, we have developed a novel fluorescence enhancement method for the determi- nation of an anionic surfactant sodium dodecyl sulfate （SDS） through the gradual optimization of experiment conditions. [ Result] Under the opti- mum conditions, the extent of fluorescence enhancement is directly proportional to SDS concentration varying from 0.5 to 16.0 μmol/L, and the de- tection limit reaches 0.051 μmol/L. The relative standard deviation （RSD） for 4.0 μmol/L SDS is 3.3% （ n =6）. The proposed method has been successfully applied to the determination of SDS in environmental water samples, with recovery of 96.3% -105.5%. E Conclusion] The novel fluo- rescence enhancement method is not only simple and rapid, but also has avoided using tedious solvent-extraction and toxic organic solvents.

A QUENCHING FLUORESCENCE IMMUNOASSAY METHOD FOR DETERMINATION OF TRACE ALBUMIN USING UNLABELED TERBIUM CHELATE

A fluoroimmunoassay method using unlabeled Terbium chelate is described.The principle is similar to that of fluoroimmunoassay method using lanthanide chelate as labels.The procedure is simpte because labeling process is unnecessary.The recovery of HSA and albumin in urine is 107% and 95% respectively.The standard deviation is tess than 10%.

Determination of Chlorobenzene by Hydroxypropyl-β- Cyclodextrin Sensitized Fluorescence Quenching Method with Neutral Red as a Fluorescence Probe

The fluorescence quenching of inclusion complex of neutral red （NR） and hydroxypropyl-β-cyclodextrin （HP-β-CD） carried by chlorobenzene was investigated. The fluorescence intensity of NR increased due to the formed inclusion complex of HP-β-CD and NR. But the fluorescence intensity of NR-HP-β-CD diminished when chlorobenzene was added, and there was a linear relationship between the fluorescence quenching value of the system （△IF = IF, NR-HP-β-CD - IF, CB-NB-NR-HP-β-CD） and the concentration of chlorobenzene. Based on this, a novel fluorescence quenching method for the determination of chlorobenzene with NR as a fluorescence probe has been developed. Under the optimal conditions, the linear range of calibration curve for the determination of chlorobenzene was 5.0 × 10^-8 - 8.0 × 10^-6 mol/L and the detection limit was 1.0 × 10^-8 mol/L. It has been applied to determination ofchlorobenzene in synthetic waste water samples with satisfactory results.

Determination of heavy metal ion in landfill leachate with solvent extracting ＆ fluorescence quenching method

Add the masking agent and biscyclohexanoneoxalyldihydraone into the diluted clarificd liquid of the landfill leachate which was treatmented by digestion and centrifugal filtration to complexate all heavy metal ion in the trcatmented liquid, and extracted using CHCl3 many time, then it were demasking and decomposing respectively, and adjusted different pH and formed Me^n＋-PAN coordination compound when these metal ion reaction with PAN. The Fluoresence quenching of Rh6G （λex/λem=543mn/558nm） when the metal ion coordination compound was add into the Rh6G solution step by step, the quenching intensity was directly proportional to the concentration of the metal ion in the certain range. So a new method of fast and simple for determination of trace metal ion in landfill lcachate was established to determine metal ion in sample of different landfill leachate in Three Gorge Water Reservoir, and comparison the classic assay method with satisfactory results.

Soller slits automatic focusing method for multi-element fluorescence detector

In X-ray absorption fine structure(XAFS) experiments,Soller slits are widely used as filter devices in order to improve the signal to noise ratio.Performing high accuracy manual focusing operations is a time-consuming process;therefore,this work introduces an automatic focusing method for Soller slits in multi-element fluorescence detectors.This method establishes a relation model between the fluorescence intensity distribution and the coordinates of the fluorescence excitation point.According to this relation model,the actual coordinates of the fluorescence excitation point can be deduced from the detected fluorescence intensity distribution and used in focusing operations.This method has proven to be feasible in an XAFS experiment at the BL14W1 beamline of the Shanghai Synchrotron Radiation Facility.

Quantitative energy-dispersive X-ray fluorescence analysis for unknown samples using full-spectrum least-squares regression

The full-spectrum least-squares(FSLS) method is introduced to perform quantitative energy-dispersive X-ray fluorescence analysis for unknown solid samples.Based on the conventional least-squares principle, this spectrum evaluation method is able to obtain the background-corrected and interference-free net peaks, which is significant for quantization analyses. A variety of analytical parameters and functions to describe the features of the fluorescence spectra of pure elements are used and established, such as the mass absorption coefficient, the Gi factor, and fundamental fluorescence formulas. The FSLS iterative program was compiled in the C language. The content of each component should reach the convergence criterion at the end of the calculations. After a basic theory analysis and experimental preparation, 13 national standard soil samples were detected using a spectrometer to test the feasibility of using the algorithm. The results show that the calculated contents of Ti, Fe, Ni, Cu, and Zn have the same changing tendency as the corresponding standard content in the 13 reference samples. Accuracies of 0.35% and 14.03% are obtained, respectively, for Fe and Ti, whose standard concentrations are 8.82% and 0.578%, respectively. However, the calculated results of trace elements (only tens of lg/g) deviate from the standard values. This may be because of measurement accuracy and mutual effects between the elements.

Tumor cell death visualization of renal cell carcinoma under the combined effect of the Gratiola officinalis extract and cyclophosphamide using fluorescent staining methods

Objective of the study:We used fluorescence imaging methods of apoptosis and necrosis in human renal carcinoma A498 tumor cells in vitro to reveal the indicated forms of cell death under the combined effect of flavonoid-containing extract of Gratiola officinalis and cytostatic(cyclo-phosphamide).Materials and methods:The dyes were propidium iodide and acridine orange,which were used in the"alive and dead"test.This test helped us to identify the total number of dead cells in the forms of necrosis and apoptosis and the number of cells in which apoptosis had started,it was characterized by the appearance of apoptotic bodies or nucleus pyknosis.Results:We found the most pronounced cytotoxic activity at the ratio of extract of Gratiola officinalis and cyclophosphamide concentrations of 1∶1.The number of living cells decreased when exposed to the ratio of extract and cytostatic concentrations of 2∶1.When the ratio of concentration of the extract relative to the cytostatic increased to 3∶1,the cytostatic activity of the extract began to appear,the total number of tumor cells decreased.The number of cells with nucleus pyknosis and the number of cells with apoptosis signs significantly increased at a 3∶1 ratio of extract and cytostatic concentrations,which confirms the presence of pro-apoptotic activity of the studied combination.This trend indicates the dependence of a certain form of cell death(apoptosis,necrosis)on the ratio of extract and cytostatic doses,and it also demon-strates the cytostatic and cytotoxic effects of this combination.Conclusion:Fluorescence methods of investigation in the"alive and dead"test allowed us to visualize the forms of cell death of human kidney carcinoma A498 by combined exposure to the fiavonoid-containing extract of Gratiola officinalis and cytostatic(cyclophosphamide)24 h after exposure.We found that the combination with a concentration ratio of the extract and cyclophosphamide of 3:1 has the greatest effectiveness due to stimulation of the cytostatic effect and cytotoxic effect.

Study on gaseous CS_2 using laser-induced fluorescence

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QM/MM Study of the Second Harmonic Generation and Two-photon Absorption Properties of a Fluorescent Proteins-Dreiklang

A new reversibly switchable fluorescent protein（RSFP）, namely Dreiklang, exhibits prominent feature that the wavelengths for switching and fluorescence are decoupled due to its great different structures between bright and dark states. This feature might also induce some nonlinear optic（NLO） properties changing as switching between two states, which might promote new method of biological science. We employ the QM/MM method to simulate the structures of different states, and study their second harmonic generation（SHG） and two-photon absorption（TPA） properties. And we found different states of Dreiklang have different SHG and TPA responses. The SHG and TPA properties of Dreiklang are correlated to particularly geometrical structures of different states, especially the centrosymmetric or nocentrosymmetric π-stacking structures which are formed by chromophore and beside residue Tyr203, so the SHG and TPA responses could be changed as the light induces switching among different states of Dreiklang. This work would prospectively guide the application of Dreiklang on the NLO technology, and help the development of new RSFP with special NLO function.

Constant Stress Accelerated Life Tests for Vacuum Fluorescent Display Based on Least Square Method

To estimate the life of vacuum fluorescent display （VFD） more accurately and reduce test time and cost, four constant stress accelerated life tests （CSALTs） were conducted on an accelerated life test model. In the model, statistical analysis of test data is achieved by applying lognormal function to describe the life distribution, and least square method （LSM） to calculate the mean value and the standard deviation of logarithm. As a result, the accelerated life equation was obtained, and then a self-developed software was developed to predict the VFD life. The data analysis results demonstrate that the VFD life submits to lognormal distribution, that the accelerated model meets the linear Arrhenius equation, and that the precise accelerated parameter makes it possible to acquire the life information of VFD within one month.