Sensitivity of diagnosis of spontaneous bacterial peritonitis is higher with the automated cell count method

BACKGROUND Spontaneous bacterial peritonitis(SBP)is one of the most important complications of patients with liver cirrhosis entailing high morbidity and mortality.Making an accurate early diagnosis of this infection is key in the outcome of these patients.The current definition of SBP is based on studies performed more than 40 years ago using a manual technique to count the number of polymorphs in ascitic fluid(AF).There is a lack of data comparing the traditional cell count method with a current automated cell counter.Moreover,current international guidelines do not mention the type of cell count method to be employed and around half of the centers still rely on the traditional manual method.AIM To compare the accuracy of polymorph count on AF to diagnose SBP between the traditional manual cell count method and a modern automated cell counter against SBP cases fulfilling gold standard criteria:Positive AF culture and signs/symptoms of peritonitis.METHODS Retrospective analysis including two cohorts:Cross-sectional(cohort 1)and case-control(cohort 2),of patients with decompensated cirrhosis and ascites.Both cell count methods were conducted simultaneously.Positive SBP cases had a pathogenic bacteria isolated on AF and signs/symptoms of peritonitis.RESULTS A total of 137 cases with 5 positive-SBP,and 85 cases with 33 positive-SBP were included in cohort 1 and 2,respectively.Positive-SBP cases had worse liver function in both cohorts.The automated method showed higher sensitivity than the manual cell count:80%vs 52%,P=0.02,in cohort 2.Both methods showed very good specificity(>95%).The best cutoff using the automated cell counter was polymorph≥0.2 cells×10^(9)/L(equivalent to 200 cells/mm^(3))in AF as it has the higher sensitivity keeping a good specificity.CONCLUSION The automated cell count method should be preferred over the manual method to diagnose SBP because of its higher sensitivity.SBP definition,using the automated method,as polymorph cell count≥0.2 cells×10^(9)/L in AF would need to be considered in patients admitted with decompensated cirrhosis.

Activity Determination of 8 Chinese Herbs against Hepatoma Cell SMMC-7721 in Vitro by MTT Method

[Objective] The aim was to build up a set of efficient and rapid models for laboratory to screen anti-hepatocellular carcinoma active substance in vitro. [ Method] By using MTT method, the activities of anti-hepatocellular carcinoma SMMC-7721 in vitro from Cymbopogon distans, Lobelia chinensis, Buddleja offlcinalis, Glycyrrhiza uralensis, Sanguisorba officinalis, Bupleurum chinense, Apium graveolen and Curuma zedoaria were tested. The growth curve of hepatoma cell was described, and the growth status in different periods were observed by inverted microscope. [ Result] Cells induced by active substance would be condensing, clear brim, which have significant differences from normal SMMC- 7721 cells. The results suggested that ESCG, ESCC, ESCB could inhibit proliferation of SMMC-7721 cells at the concentration of 1.0 -1.5 mg/ml, and the inhibition rate were 51.6%, 48.5%, 52.9% respectively. With the increasing of concentration, the inhibition strengthened. [ Conclusion] MTT method could be used as a basic model for screening important anti-hepatoma.

Highly efficient solution-processed CZTSSe solar cells based on a convenient sodium-incorporated post-treatment method

In CZTSSe solar cells,a simple sodium-incorporation post-treatment method toward solution-processed Cu2Zn Sn S4precursor films is presented in this work.An ultrathin NaCl film is deposited on Cu2Zn Sn S4precursor films by spin-coating NaCl solution.In subsequent selenization process,the introduction of Na Cl is found to be benefacial for the formation of Cu2-xSe,which can further facilitate the element transportation,leading to dense and smooth CZTSSe films with large grains and less impurity Cu2Sn(S,Se)3phase.SIMS depth profiles confirm the gradient distribution of the sodium element in Na-doped absorbers.Photoluminescence spectra show that the introduction of appropriate sodium into the absorber can inhibit the band tail states.As high as 11.18% of power conversion efficiency(PCE)is achieved for the device treated with 5 mg mL^-1 NaCl solution,and an average efficiency of Na-doped devices is 10.71%,13%higher than that of the control groups(9.45%).Besides,the depletion width and the charge recombination lifetime can also have regular variation with sodium treatment.This work offers an easy modification method for high-quality Na-doped CZTSSe films and high-performance devices,in the meantime,it can also help to further understand the effects of sodium in CZTSSe solar cells.

PREDICTION OF TEXTILE FABRIC REINFORCED COMPOSITE PROPERTIES BASED ON NODE INTERPOLATION CELL METHOD

Node interpolation cell method（NICM）is a micromechanics method employing the virtual displacement principle and the representative volume element（RVE）scheme to obtain the relationship between the global and the local strain.Mechanical properties of 2-D textile fabric reinforced ceramic matrix composites are predicted by NICM.Microstructures of 2-D woven and braided fabric reinforced composite are modeled by two kinds of RVE scheme.NICM is used to predict the macroscopic mechanical properties.The fill and warp yarns are simulated with cubic B-spline and their undulating forms are approximated by sinusoid.The effect of porosity on the fiber and matrix are considered as a reduction of elastic module.The connection of microstructure parameters and fiber volume fraction is modeled to investigate the reflection on the mechanical properties.The results predicted by NICM are compared with that by the finite element method（FEM）.The comparison shows that NICM is a valid and feasible method for predicting the mechanics properties of 2-D woven and braided fabric reinforced ceramic matrix composites.

Finite cell method compared to h-version finite element method for elasto-plastic problems

The finite cell method （FCM） combines the high-order finite element method （FEM） with the fictitious domain approach for the purpose of simple meshing. In the present study, the FCM is used to the Prandtl-Reuss flow theory of plasticity, and the results are compared with the h-version finite element method （h-FEM）. The numerical results show that the FCM is more efficient compared to the h-FEM for elasto-plastic problems, although the mesh does not conform to the boundary. It is also demonstrated that the FCM performs well for elasto-plastic loading and unloading.

ONE-CELL-STATE METHOD FOR DETERMINATION OF ELECTRONIC STRUCTURE OF INTERMETALLIC COMPOUNDS

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Methods to produce induced pluripotent stem cell-derived mesenchymal stem cells: Mesenchymal stem cells from induced pluripotent stem cells

Mesenchymal stem cells(MSCs)have received significant attention in recent years due to their large potential for cell therapy.Indeed,they secrete a wide variety of immunomodulatory factors of interest for the treatment of immune-related disorders and inflammatory diseases.MSCs can be extracted from multiple tissues of the human body.However,several factors may restrict their use for clinical applications:the requirement of invasive procedures for their isolation,their limited numbers,and their heterogeneity according to the tissue of origin or donor.In addition,MSCs often present early signs of replicative senescence limiting their expansion in vitro,and their therapeutic capacity in vivo.Due to the clinical potential of MSCs,a considerable number of methods to differentiate induced pluripotent stem cells(iPSCs)into MSCs have emerged.iPSCs represent a new reliable,unlimited source to generate MSCs(MSCs derived from iPSC,iMSCs)from homogeneous and well-characterized cell lines,which would relieve many of the above mentioned technical and biological limitations.Additionally,the use of iPSCs prevents some of the ethical concerns surrounding the use of human embryonic stem cells.In this review,we analyze the main current protocols used to differentiate human iPSCs into MSCs,which we classify into five different categories:MSC Switch,Embryoid Body Formation,Specific Differentiation,Pathway Inhibitor,and Platelet Lysate.We also evaluate common and method-specific culture components and provide a list of positive and negative markers for MSC characterization.Further guidance on material requirements to produce iMSCs with these methods and on the phenotypic features of the iMSCs obtained is added.The information may help researchers identify protocol options to design and/or refine standardized procedures for large-scale production of iMSCs fitting clinical demands.

Study on the packed volume and the void ratio of idealized human red blood cells using a ?nite-discrete element method

Numerical simulations are performed to examine the packing behavior of human red blood cells(RBCs). A combined ?nite-discrete element method(FDEM) is utilized, in which the RBCs are modeled as no-friction and no-adhesion solid bodies. The packed volume and the void ratio of a large number of randomly packed RBCs are clari?ed,and the effects of the RBC shape, the mesh size, the cell number, and the container size are investigated. The results show that the packed human RBCs with normal shape have a void ratio of 28.45%, which is slightly higher than that of the ?at or thick cells used in this study. Such information is bene?cial to the further understanding on the geometric features of human RBCs and the research on RBC simulations.

Preparation of Se-based solar cell using spin-coating method in ambient condition

A new hybrid organic-inorganic structure of FTO/TiO2/Se/HTL/Au based selenium solar cell has been fabricated through a low-cost spin-coating process in air. In this process, selenium is completely dissolved in hydrazine, to fk）rm a homogeneous precursor solution. After spin-coating the precursor solution on the TiO2 substrates, following by sintering at 200 ℃ for 5rain, a uniform selenium film with crystalline grains is formed. The selenium based solar cell exhibits an efficiency of 1.23% under AM1.5 illumination （100 mW.cm-2）, short-circuit current density of 8 mA.cm 2, open-circuit voltage of 0.55 V, and fill factor of 0.37. Moreover, the device shows a stable ability with almost the same performance alter 60 days.

MESHLESS METHOD FOR 2D MIXED-MODE CRACK PROPAGATION BASED ON VORONOI CELL

A meshless method integrated with linear elastic fracture mechanics(LEFM)is presented for 2D mixed-mode crack propagation analysis.The domain is divided automatically into sub-domains based on Voronoi cells,which are used for quadrature for the potential energy. The continuous crack propagation is simulated with an incremental crack-extension method which assumes a piecewise linear discretization of the unknown crack path.For each increment of the crack extension,the meshless method is applied to carry out a stress analysis of the cracked structure.The J-integral,which can be decomposed into mode Ⅰ and mode Ⅱ for mixed-mode crack,is used for the evaluation of the stress intensity factors(SIFs).The crack-propagation direction,predicted on an incremental basis, is computed by a criterion defined in terms of the SIFs. The flowchart of the proposed procedure is presented and two numerical problems are analyzed with this method.The meshless results agree well with the experimental ones,which validates the accuracy and efficiency of the method.

Preparation of CH_3NH_3PbI_3 thin films for solar cells via Vapor Transfer Method

Organometal halide perovskite based solar cells have emerged as one of the most promising candidates for low-cost and high-efficiency solar cell technologies. Here a Vapor Transfer Method （VTM） is used to fabricate high quality perovskite thin films in a balanced vacuum capsule. By adjusting the reaction tem- perature, CH_3NHl_3 saturated vapor which then reacts with Pbl_2 films can be controlled and the formation process of CH_3NH_3Pbl_3 perovskite films can be further influenced. Prepared perovskite films which ex- hibit pure phase, smooth surface and high crystallinity are assembled into planar heterojunction inverted solar cells. The whole fabrication process of solar cell devices is organic solution free. Finally, the cham- pion cell achieved power conversion efficiency （PCE） of 13.08% with negligible current-voltage hysteresis under fully open-air conditions. The photovoltaic performance could be further enhanced by optimizing perovskite composition and the device structure.

Shape optimization of axisymmetric solids with the finite cell method using a fixed grid

In this work, a design procedure extending the B-spline based finite cell method into shape optimization is developed for axisymmetric solids involving the centrifugal force effect. We first replace the traditional conforming mesh in the finite element method with structured cells that are fixed during the whole design process with a view to avoid the sophisticated re-meshing and eventual mesh distortion.Then, B-spline shape functions are further implemented to yield a high-order continuity field along the cell boundary in stress analysis. By means of the implicit description of the shape boundary, stress sensitivity is analytically derived with respect to shape design variables. Finally, we illustrate the efficiency and accuracy of the proposed protocol by several numerical test cases as well as a whole design procedure carried out on an aeronautic turbine disk.

On-line Measurement for Ohmic Resistance in Direct Methanol Fuel Cell by Current Interruption Method

Electrochemical impedance spectroscopy (EIS) is widely used in fuel cell impedance analysis. However, for ohmic resistance (R Ω), EIS has some disadvantages such as long test period and complex data analysis with equivalent circuits. Therefore, the current interruption method is explored to measure the value of RΩ in direct methanol fuel cells (DMFC) at different temperatures and current densities. It is found that RΩ decreases as temperature increase, and decreases initially and then increases as current density increases. These results are consistent with those measured by the EIS technique. In most cases, the ohmic resistances with current interruption (R iR ) are larger than those with EIS (R EIS ), but the difference is small, in the range from –0.848% to 5.337%. The errors of R iR at high current densities are less than those of R EIS . Our results show that the R iR data are reliable and easy to obtain in the measurement of ohmic resistance in DMFC.

Generation of Transgene-free Induced Pluripotent Stem Cells with Non-viral Methods

Induced pluripotent stem （iPS） cells were originally generated from mouse fibroblasts by enforced expression of Yamanaka factors （Oct3/4, Sox2, Klf4, and c-Myc）. The technique was quickly re- produced with human fibroblasts or mesenchymal stem cells. Although having been showed therapeutic po- tential in animal models of sickle ceil anemia and Parkinson＇s disease, iPS cells generated by viral methods do not suit all the clinical applications. Various non-viral methods have appeared in recent years for application of iPS cells in cell transplantation therapy. These methods mainly include DNA vector-based approaches, transfection of mRNA, and transduction of reprogramming proteins. This review summarized these non-viral methods and compare the advantages, disadvantages, efficiency, and safety of these methods.

Computational Study of Collective Cell Migration By Meshfree Method

The collective cell migration behavior on a substrate was studied using RKPM meshfree method.The cells were modeled as nematic liquid crystal with hyperelastic cell nucleus.The cell-substrate and cell-cell interactions were modeled by coarse-grained potential forces.Through this study,the pulling and pushing phenomenon during collective cell migration process was observed and it was found that the individual cell mobility significantly influenced the collective cell migratory behavior.More self-propelled cells are in the system along the same direction,the faster the collective group migrates toward coordinated direction.The parametric study on cell-cell adhesion strength indicated that as the adhesion strength increases,the collective cell migration speed increases.It also showed that the mechanical stress in leader cell is higher than stress in follower cells.

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.

Establishment and validation of a method for cell irradiation in 96-well and 6-well plates using a linear accelerator

To establish and validate a method for cell irradiation in 96-well and 6-well plates using a linear accelerator, three irradiation methods(G0 B0 F40,G0 B1.5 F40, and G180 B1.5 F40) were designed to irradiate cell culture plasticware simulated with RW3 slab phantom and polystyrene. The difference between the actual physical measured dose and the preset dose was compared among the three methods under the preparatory conditions of 2, 4, 6, 8, and 10 Gy. MDA-MB-231 cells were analyzed by using a cell proliferation assay and a clonogenic assay to verify the difference between the three cell irradiation methods on cell radiosensitivity. For each preset dose, the difference between the actual measured dose and the preset dose was the lowest for Method G0 B1.5 F40, the second lowest for Method G180 B1.5 F40, and the maximum for Method GOB0 F40. The ranges of the differences were-0.28 to 0.02%,-2.17 to-1.80%, and-4.92 to-4.55%, and 0.31 to-0.12%,-3.42 to-2.86%, and-7.31 to-6.92%,respectively, for 96-well and 6-well plates. The cell culture experiments proved that Method G0 B1.5 F40 was an accurate, effective, simple, and practical irradiation method. The most accurate and effective cell irradiation method should always be used, as it will reduce dose differences and instability factors and provide improved accuracy and comparability for laboratories researching cellular radiosensitivity.

In vitro cultivation of rat bone marrow mesenchymal stem cells and establishment of pEGFP/Ang-1 transfection method

Objective:To obtain the bone marrow mesenchymal stem cells(BMSCs).complete phenotypic identification and successfully transfecl rat BMSCs by recombinant plasmid pF.GFP/Ang-1.Methods:BMSCs were isolated from bone marrow using density gradient centrifugation method and adherence screening method,and purified.Then the recombinant plasmid pEGFP/Ang-1was used to transfect BMSCs and the positive clones were obtained by the screen of C418 and observed under light microscopy inversely.Green fluorescent exhibited by protein was enhanced to measure the change time of the expression amount of Ang-1.Results:BMSCs cell lines were obtained successfully by adherence screening method and density gradient ccntrifugation.Ang-1 recombinant plasmid was transfected smoothly into rat BMSCs,which can express Ang-1 for 3 d and decreased after 7 d.Conclusions:Adherence screening method und density gradient ceiilrifugation can be effective methods lo obtain BMSCs with high purity and rapid proliferation.Besides,the expression of transfected recombinant plasmid pEGFP/Ang-1 in rat BMSCs is satisfactory.

A porous cell method for the simulation of fluid-solid interactions

There are many ways of describing a solid,porous or fluid region of the computational domain when solving the Navier-Stokes equations(NSE)for flow motions.Amongst these the porous cell method is one of the most flexible approaches.In this method,a parameter is defined as a ratio of the volume open to water and air in a calculation cell to its cell volume.In the calculation,the same numerical procedure is applied to every cell and no explicit boundary conditions are needed at solid boundaries.The method is used to simulate flow through porous media,around solid bodies and over a moving seabed.The results compare well with experimental data and other numerical results.In our future work the porous cell method will be applied to more complex fluid-solid interaction situations.

Study of a Quantitative Fluorescence Method for Interleukin 2 Receptor of Mononuclear Cell in Normal Human Peripheral Blood

A quantitative method using fluorescence spectro-photometer to detect theinterleukin 2 reocptor （IL 2R） on the surface of mononuclear cells in normal humanperipheral blood is described. For expression of IL 2R, the optimum conoentration of PHA andCon A were 100 μg/ml and 10 μg/ml respectively. The PHA induced effect on mononuclear cellswas better than Con A induced one. The peak value of IL 2R cxpression was at the 24th h afterstimulation The optimum dilution of the first antibody （anti-Tac） was 1: 50, while the dilution ofthe second antibody（fluorescein conjugated rabbit anti-mousc IgG） was 1: 32 Their optimumincubation period was 20 and 10 min repectively. The concentration of fluorcscein-conjugatedantibody per 1×106 mononuclear cells from the same normal human peripheral blood was5.22±0.45×10-10 mol, and the coefficient of variation was 8. 6%. The concentration offluorescein-conjugated antibody per 1×106PBMC from five healthy individuals was 4. 8±0. 97×10-10 mol Therefore this assay system is stable and quite reproducible.