Down-regulation of Hsp90 could change cell cycle distribution and increase drug sensitivity of tumor cells

AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal cancer cell lines, and to study the cell cycle distribution of the gene transected cells and their response to chemotherapeutic drugs.METHODS:A 1.03kb cDNA sequence of Hsp90beta was obtained from the primary plasmid phHSP90 by EcoR I and BamH I nuclease digestion and was cloned to the EcoR I and BamH I site of the pcDNA by T4DNA ligase and an antisense orientation of Hsp90beta expression vector was constructed. The constructs were transfected with lipofectamine and positive clones were selected with G418. The expression of RNA was determined with dot blotting and RNase protection assay, and the expression of Hsp90 protein determined with western blot. Cell cycle distribution of the transfectants was analyzed with flow cytometry, and the drug sensitivity of the transfectants to Adriamycin (ADR), vincrinstine (VCR), mitomycin (MMC) and cyclophosphamide (CTX) with MTT and intracellular drug concentration of the transfectants was determined with flow cytometry.RESULTS:In EcoR I and BamH I restriction analysis, the size and the direction of the cloned sequence of Hsp90beta remained what had been designed and the gene constructs were named pcDNA-Hsp90.AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones all expressed Hsp90 anti-sense RNA. The expression of Hsp90 was down-regulated in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones. Cell cycle distribution was changed differently. In AH-SGC7901/VCR and AH-Ec109 cells, G(1) phase cells were increased; S phase and G(2) phase cells were decreased as compared with their parental cell lines. In AH-SGC7901 cell, G(1)phase cells were decreased, G(2) phase cells increased and S phase cells were not changed, and in AH-HCC7402 cells G(1), S and G(2) phase cells remained unchanged as compared with their parental cell lines. The sensitivity of AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 to chemotherapeutic drugs, the sensitivity of AH-SGC7901/VCR to ADR, VCR, MMC and CTX the sensitivity of AH-HCC7402 to ADR and VCR, and the sensitivity of Ec109 to ADR, VCR and CTX all increased as compared with their parental cell lines. The mean fluorescence intensity of ADR in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 was also significantly elevated (P 【 0.05).CONCLUSION: Down-regulation of Hsp90 could change cell cycle distribution and increase the drug sensitivity of tumor cells.

Effects of hypoxia on promoter of telomerase reverse transcriptase and cell cycle distribution in neonatal rat cardiac myocytes

On the hypothesis that telomerase reverse transcriptase (TERT) of cardiac myocytes (CMs) is consistent with cell cycle distribution as well as tumour cells, we plan to investigate the expression of TERT in CMs and how TERT is in keeping with CMs cycle distribution after birth and under hypoxia, and roughly understand how hypoxia affects activity of TERT promoter.

Temperature field distribution of burnt surrounding rock in UCG stope

In order to study temperature field distribution in burnt surrounding rock and to determine ranges of burnt surrounding rock, coal-wall coking cycle and heat influence in the underground coal gasification(UCG) stope, based on the Laplace transform and inversion formula, we studied the temperature analytical solution of one-dimensional unsteady heat conduction for multi-layer overlying strata under the first and the forth kinds of boundary conditions, and we also carried out a numerical simulation of twodimensional unsteady heat conduction by the COMSOL multiphysics. The results show that when the boundary temperature of surrounding rock has a linear decrease because of a directional movement of heat source in the UCG flame working face, the temperature in surrounding rock increases first and then decreases with time, the peak of temperature curve decreases gradually and its position moves inside surrounding rock from the boundary. In the surrounding rock of UCG stope, there is an envelope curve of temperature curve clusters. We analyzed the influence of thermophysical parameters on envelope curves and put forward to take envelope curve as the calculation basis for ranges of burnt surrounding rock, coal-wall coking cycle and heat influence. Finally, the concrete numerical values are given by determining those judgement standards and temperature thresholds, which basically tally with the field geophysical prospecting results.

Probabilistic Analysis and Fatigue Damage Assessment of Offshore Mooring System due to Non-Gaussian Bimodal Tension Processes

Both wave-frequency(WF) and low-frequency(LF) components of mooring tension are in principle non-Gaussian due to nonlinearities in the dynamic system.This paper conducts a comprehensive investigation of applicable probability density functions(PDFs) of mooring tension amplitudes used to assess mooring-line fatigue damage via the spectral method.Short-term statistical characteristics of mooring-line tension responses are firstly investigated,in which the discrepancy arising from Gaussian approximation is revealed by comparing kurtosis and skewness coefficients.Several distribution functions based on present analytical spectral methods are selected to express the statistical distribution of the mooring-line tension amplitudes.Results indicate that the Gamma-type distribution and a linear combination of Dirlik and Tovo-Benasciutti formulas are suitable for separate WF and LF mooring tension components.A novel parametric method based on nonlinear transformations and stochastic optimization is then proposed to increase the effectiveness of mooring-line fatigue assessment due to non-Gaussian bimodal tension responses.Using time domain simulation as a benchmark,its accuracy is further validated using a numerical case study of a moored semi-submersible platform.

On Simple MCD-Graphs

A simple graph G on n vettices is said to be a simple MCD-graph if G has no two cyties having the same length and has the maximum possible number of edges.Two results of the number of cy cles in G are given by introdueing the Concept of a path decomposition and by them,the following theorem is proved:If G is a simple MCD-graph,then G is not a 2-connected planar graph and for all n except seven integer,G is not a 2-connected graph on n vertices containing a subgraph homeomor phic to K_4.

Influence of cycle time distribution on coating uniformity of particles in a spray fluidized bed by using CFD-DEM simulations

Cycle Time Distribution(CTD)plays a critical role for determining uniformity of particle coating in spray fluidized beds.However,the CTD is influenced by both geometrical structure and operating conditions of fluidized bed.In this study,a spray fluidized bed of coating process is simulated by a comprehensive Computational Fluid Dynamics-Discrete Element Model(CFD-DEM).To achieve different behaviors of CTD,some modifications are designed on a pseudo-2D internally circulating fluidized bed,which traditionally composes of a high-velocity upward bed and low-velocity downward bed.These modifi-cations include making the air distributor slope and/or laying a baffle in the downward bed.First,the CTD and evolution of particle size distribution under different bed structures are compared.The CTD directly influences the coating uniformity.By making the particles flowing along a parallel direction in the downward bed through the geometrical modifications,the CTD becomes narrower and the coating uniformity is significantly improved.Second,under the optimized bed structure,the influence of oper-ating conditions on the coating uniformity is studied.Properly increasing the fluidization gas velocity and the fluidization gas temperature and reducing the liquid spray rate can improve the coating uniformity.

Uniqueness of Cycle Length Distributions of Certain Bipartite Graphs K_(n,n+7)-A(|A| ≤ 3)

The cycle length distribution of a graph of order n is denoted by (c1,c2,...,cn), where ci is the number of cycles of length i. In this paper, we obtain that a graph G is uniquely determined by its cycle distribution if: (1) G = Kn,n+7 (n ≥ 10); or (2) G = Kn,n+7 - A (|A| = 1,n ≥ 12); or (3) G = Kn,n+7 - A (|A| = 2,n ≥ 14); or (4) G = Kn,n+7 - (|A| = 3,n ≥ 16), where A - E(Kn,n+7).

Bipartite Graphs K_(n,n+r)-A(|A|≤3) Determined by Their Cycle Length Distributions

The cycle length distribution of a graph G of order n is a sequence （c1 （G）,…, cn （G））, where ci （G） is the number of cycles of length i in G. In general, the graphs with cycle length distribution （c1（G） ,…,cn（G）） are not unique. A graph G is determined by its cycle length distribution if the graph with cycle length distribution （c1 （G）,…, cn （G）） is unique. Let Kn,n＋r be a complete bipartite graph and A lohtaib in E（Kn,n＋r）. In this paper, we obtain： Let s 〉 1 be an integer. （1） If r = 2s, n 〉 s（s - 1） ＋ 2｜A｜, then Kn,n＋r - A （A lohtain in E（Kn,n＋r）,｜A｜ ≤ 3） is determined by its cycle length distribution; （2） If r = 2s ＋ 1,n 〉 s^2 ＋ 2｜A｜, Kn,n＋r - A （A lohtain in E（Kn,n＋r）, ｜A｜ ≤3） is determined by its cycle length distribution.

DISTRIBUTIONS OF LIMIT CYCLES FOR POLYNOMIAL VECTOR FIELDS (P_2, Q_3)

The distributions of limit cycles of cubic vector fields (P2, Q3) are considered in this paper, where P2 and Q3 are polynomials of x and y of order two and three, respectively. It is possibly seven different distributions of limit cycles given in [1]. We now prove that in which three kinds of distributions are impossible and other four kinds all can be realized by concrete vector fields of (P2,Q3). Some related results are also given.

Bifurcations of Limit Cycles in a Z_4-Equivariant Quintic Planar Vector Field

In this paper, a Z4-equivariant quintic planar vector field is studied. The Hopf bifurcation method and polycycle bifurcation method are combined to study the limit cycles bifurcated from the compounded cycle with 4 hyperbolic saddle points. It is found that this special quintic planar polynomial system has at least four large limit cycles which surround all singular points. By applying the double homoclinic loops bifurcation method and Hopf bifurcation method, we conclude that 28 limit cycles with two different configurations exist in this special planar polynomial system. The results acquired in this paper are useful for studying the weakened 16th Hilbert＇s Problem.