The enhanced genomic 6 mA metabolism contributes to the proliferation and migration of TSCC cells / 国际口腔科学杂志·英文版

In contrast to the well-established genomic 5-methylcytosine (5mC), the existence of N6-methyladenine (6 mA) in eukaryotic genomes was discovered only recently. Initial studies found that it was actively regulated in cancer cells, suggesting its involvement in the process of carcinogenesis. However, the contribution of 6 mA in tongue squamous cell carcinoma (TSCC) still remains uncharacterized. In this study, a pan-cancer type analysis was first performed, which revealed enhanced 6 mA metabolism in diverse cancer types. The study was then focused on the regulation of 6 mA metabolism, as well as its effects on TSCC cells. To these aspects, genome 6 mA level was found greatly increased in TSCC tissues and cultured cells. By knocking down 6 mA methylases N6AMT1 and METTL4, the level of genomic 6 mA was decreased in TSCC cells. This led to suppressed colony formation and cell migration. By contrast, knockdown of 6 mA demethylase ALKBH1 resulted in an increased 6 mA level, enhanced colony formation, and cell migration. Further study suggested that regulation of the NF-κB pathway might contribute to the enhanced migration of TSCC cells. Therefore, in the case of TSCC, we have shown that genomic 6 mA modification is involved in the proliferation and migration of cancer cells.

Effects and the mechanisms of cardiac short-term memory on cellular electrical excitability / 生物医学工程学杂志

Electrical instability easily induces a unidirectional conduction block, resulting in ventricular tachycardia (VT) or even fibrillation (VF). Cardiac memory affects dynamic electrical characteristics through previous pacing so that it makes the memory important in arrhythmia study. This paper investigates the impact of the rapid pacing duration on cellular excitability and its mechanism. Based on the canine endocardial single cell, a one-dimensional tissue model was developed. Simulations were realized with OpenMP parallel programming method. The results showed that with repetitive pacing, the cellular excitability became low while the conduction velocity decreased. Accumulation of intracellular [Ca2+]i and [Na+]i and depletion of [K+]i led to the shift of membrane current-voltage curves, changing the membrane resistance. Excitability determined by the resistance at the large width of stimulus pulse, therefore, it suggested that [Ca2+]i and [K+]i-induced memory formed the ionic substrates for the alteration of excitability.

Algorithm study on the three-dimensional cardiac tissue based on the model of ventricular action potential / 生物医学工程学杂志

Cardiac reentry is one of the important factors to induce arrhythmias. It could lead to ventricular tachycardia (VT) or even fibrillation (VF), resulting in sudden cardiac death. With the wide use of computer in the quantitative study of electrophysiology, the three-dimensional virtual heart for simulations needs to be developed imminently in computer. In this paper, numerical algorithm of the model was studied. The three-dimensional model was constructed by integrating Luo-Rudy 1991 ventricular cell model and diffusion equation. The operator splitting method was employed to solve the model. The alternate direction iterative (ADI) format and seven-point centered difference method were used for the partial differential equation. And the discrete format with second-order accuracy was taken for the boundary conditions. The results showed that the ADI format and seven-point centered difference method both could successfully figure out the membrane potential and electrical activities with good numerical stability. However, computing consumption could be greatly reduced with the ADI format, implying that the ADI method with large time step was more powerful in numerical simulations.

Toxicity to the normal hemocytes by ALA-PDT for the ex vivo purging of hematopoietic stem cell grafts / 药物分析学报

Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control;Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change;however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA,except for PBMCs, intraceUuiar PplX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosts. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologons bone marrow or stem cell grafts.

Two-photon Fluorescence Imaging of Living Cells' Anionic Sites Labeled by Cationic Colloidal Gold and Its Nano-scale Microeffect / 生物化学与生物物理进展

Cationic colloidal gold (CCG) nanoparticles were used for labeling on the anioinic sites of living cells under two-photon fluorescence (TPF) microscope,and for delivering macromolecules into the target cells when irradiated by focused femtosecond laser pulses. 15 nm CCG nanoparticles which were made by conjugation with poly-L-Lysine,were attached on the anionic sites,especially on the membrane,of CHO-K1 cells because of their strong positive charge at physiological pH. Target cells labeled with cationic gold nanoparticles were imaged under TPF microscope,and lifetime images of the same targets were taken by time correlated single photon counting (TCSPC) technique in order to verify the fluorescence of the marker and the luminescence of the gold particles. The results shown that CCG nanoparticles first accumulated on the negatively charged sites of the membrane,then entered via endocytic pathway and attached anionic sites in plasma. A macromolecular 10 ku fluorescein isothiocyanate dextran (FITC-D) was added into the sample and the focused femtosecond laser of TPL microscope was employed to scan the target cells layer by layer. Typical laser power level used in biological imaging is about 3～5 mW. Here the laser power of scanning was below 5 mW in order to prevent photochemical damage of the fs-pulses alone and to localize effects to the nanoparticles on a nano-scale. After scanning the target cells under stack mode,macromolecular fluoresceins surrounding the cells was observed to cross the membrane and to diffuse in the cytoplasma. Comparing with the images before scanning,the two-photon fluorescence and fluorescence lifetime images revealed the delivery of FITC-D into target cells. Photothermal effects,which may be responsible for the permeabilisation,are highly localized in nanoscale and are not expected to cause damage exceeding the cell membrane. After extensive of laser scanning also cell death occurred. The ratio of the uptake of FITC-D and cellular death under different conditions were measured by flow cytometer. The results shown: with the increased scanning times or ratio of particles to cells,transfer efficiency increased first and decreased afterwards,but the ratio of cellular death went up all along.

Computer simulation study of the re-entry mechanisms in one-dimensional ischaemic myocardium / 生物医学工程学杂志

Torsades de Pointes is a kind of severe ventricular arrhythmia. Myocardial ischaemia is one of the major causes leading to TdP. In this paper the mechanisms of the TdP were quantitatively studied under the condition of ischaemia based on the Noble98 dynamic model of the ventricular action potential. The study was conducted on one-dimensional homogeneous myocardium with the method of computer simulation. The models were firstly developed to simulate the lower excitability, extracellular accumulation of the K+ concentration or the decreased gap junctions in ischaemic myocardium. By separately reducing the Na+ conductance, increasing the extracellular K+ concentration or decreasing the conductance of the gap junctions enabled us to study the effect of each change in isolation. Then different degrees of ischaemic models were established to study their physiological features. The study showed that the conduction velocity became slower with the ischaemia aggravation, the action potential duration became shorter and the width of the vulnerable window obviously became larger than the normal conditions. The results illustrated that ischaemia was easily leading to unidirectional conduction block and resulted in re-entry and arrhythmias.

Study of cellular electrophysiology based on Noble98 dynamic model of ventricular action potential / 生物医学工程学杂志

On the basis of mammalian ventricular action potential model Noble98, and with the use of Runge-Kutta for solution, the Wenckebach periodicity phenomenon, the transmural heterogeneity of the ventricular myocardium and its rate dependence are studied. The results indicate that these inherent properties may, lead to temporal-space disorganized in the normal heart,and may become the underlying factors for arrhythmias. At the same time, in this study are established the basic methods for quantitative cellular electrophysiology which is essential for future studies on the mechanism of arrhythmia.

Optical mapping of the membrane potential with voltage-sensitive dyes / 生物医学工程学杂志

Optical mapping of the membrane potential with voltage-sensitive dyes is an advanced approach that involves in many theories such as molecular photonica, physiology and computer science. Compared with the conventional techniques of membrane patch-clamp and microelectrode, the optical mapping system can measure not only the changes of a single membrane potential from multiple sites simultaneously but also the conduction properties of the cell populations. So this technique provides an important method for studying the electrophysiology of the small cell, the tiny neurite and the cardiac arrhythmia, etc. Because this technique can also avoid the electromagnetic interferences, it uniquely provides an ideal means for studying the mechanisms such as cardiac defibrillation. In this paper the principles and the system structure of the optical mapping are introduced, its applications and future developments are also presented at the same time.

OPTIMIZATIONS FOR 5-AMINOLEVULINIC ACID BASED PHOTODYNAMIC THERAPY IN PURGING LEUKEMIA CELL HL60 / 药物分析学报

Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). Methods ALA incubation time, wavelength applied to irradiate, concentration of ALA incubated, irradiation fluence may modulate the effect of 5-aminolevulinic acid based Photodynamic Therapy (ALA-PDT).The high-pressure mercury lamps of 400W served as light source, the interference filter of 410nm, 432nm, 545nm, 577nm were used to select the specific wavelength. Fluorescence microscope was used to detect the fluorescence intensity and location of protoporphyrin IX (PpIX) endogenously produced by ALA. MTT assay was used to measure the survival of cell. Flow cytometry with ANNEXIN V FITC kit (contains annexin V FITC, binding buffer and PI) was used to detect the mode of cell death. Results ① 1mmol/L ALA incubated 1×105/mL HL60 cell line for 4 hours, the maximum fluorescence of ALA induced PpIX was detected in cytomembrane. ② Irradiated with 410nm for 14.4J/cm2 can result in the minimum survivability of HL60 cell. ③ The main mode of HL60 cell death caused by ALA-PDT is necrosis. Conclusion ALA for 1mmol/L, 4 hours for dark incubation time, 410nm for irradiation wavelength, 14.4J/cm2 for irradiation fluence were the optimal parameters to selectively eliminate promyelocytic leukemia cell HL60 by ALA based PDT. The photosensitization of ALA based PDT caused the necrosis of HL60 cell, so it could be used for inactivation of certain leukemia cells.

Study of Reentrant Waves in Simulated Myocardial Ischemia / 航天医学与医学工程

Objective To study ischemic effects on reentrant activities and cardiac arrhythmias using a computational approach. Method The Noble 98 mathematical model of ventricular cell was used in the study. The operator splitting and adaptive time step methods were utilized to integrate the partial differential equations in cardiac conduction models. The ischemic cells were simulated by decreasing the intracellular ATP concentration, reducing the Na+ conductance, and increasing the extracellular K+ concentration in a two-dimensional tissue. Spiral waves were initiated by the cross field technique. Result The results showed that spiral waves in local severe ischemia displayed three different morphologies,whereas in moderate ischemia only two kinds of wave forms exhibited. When the degree of ischemia reached a critical value, the reentrant wave could break. But for larger areas of ischemia spiral wave formed a typical functional reentry around the obstacle. Conclusion The study demonstrates that size and level of ischemia have effects on VTs and VFs. Large ischemic area is beneficial for maintenance of spiral wave and can provide a high probability in the genesis of VTs. Spiral waves can easily break up and degenerate into VFs under critical area or level of ischemia.

An experimental study on destruction of K562 and HL60 induced by 5-aminolaevulinic acid-based photodynamic therapy / 生物医学工程学杂志

This experiment was designed to explore the pattern of K562 and HL60 leukemia cells death, the effects on their cell cycle and the cytoplasmic free calcium concentration ([Ca2+]i) induced by 5-aminolaevulinic acid-based photodynamic therapy (ALA-PDT). Under the transmission electron microscope (TEM), two kinds of leukemia cells' ultrastructure were observed. Flow cytometry combined with Annexin V-FITC/PI labeling was used to detect the pattern of K562 and HL60 cells' death induced by ALA-PDT. Flow cytometry combined with PI labeling was used to analyze the change in the cell cycle induced by ALA-PDT, and confocal laser scanning microscopy (CLSM) combining with calcium fluorescence probe was used to detect the change in the cytoplasmic free calcium concentration ([Ca2+]i). Immediately after irradiation, many typical apoptotic bodies were seen in the cells treated. Most of the cells treated were necrotic at 24 hours following irradiation. Flow cytometry analysis suggested that the main patterns of the cells' death were apoptosis immediately after irradiation and necrosis post-apoptosis at 24 hours post irradiation. Immediately and 24 hours after irradiation, the proportion of S phase of K562 was 57. 67% +/- 1.13% and 84.77% +/- 6.20% respectively, and the proportion of S phase of HL60 was 74.60% +/- 7.27% and 84.60% 1.74% respectively. Both [Ca+]i of the treated K562 and HL60 were increased obviously. In the best experiment condition, the initial pattern of the K562 and HL60 leukemia cells' death induced by PDT was apoptosis and the main pattern was necrosis post apoptosis. The two kinds of cells were arrested at S phase by ALA-PDT. During the death of the leukemia cells, the increase in intracellular free calcium concentration could be responsible for the ALA photodynamically induced damage to K562 and HL60 cells.

Mathematical model of cardiac action potential and its computer simulations / 生物医学工程学杂志

Malignant arrhythmias and ventricular fibrillation are generally accepted as one of the major causes of death in cardiovascular diseases. Based on the H-H equations, the mathematical model of the cardiac cell action potential consists of the ion channels, pumps, exchangers and transporters that are closely connected with intra- and extra-cellular ion concentrations, the channel's conditions, nerve transductors and drugs. It can build the link between cell electrophysiology and clinical pathophysiology. By altering the cellular environments the computer simulating study on this kind of model can help us look into the electrophysiological changes of the cardiac tissue and even the whole heart and investigate the mechanisms of the cardiac arrhythmias as well. The components of the model and its computer simulating study are introduced in the paper.

Synthesis and characteristics of porous hydroxyapatite bioceramics / 生物医学工程学杂志

The macroporous structure of human bone allows the ingrowth of the soft tissues and organic cells into the bone matrix, profits the development and metabolism of bone tissue, and adapts the bone to the change of load. There is great requirement for artificial biomimic porous bioactive ceramics with the similar structure of bone tissue that can be used clinically for repairing lost bone. Fine hydroxyapatite (HAp) powder produced by wet chemical reaction was mixed with hydrogen peroxide (H2O2), polyvinyl alcohol, methyl cellulose or other pores-making materials to form green cake. After drying at low temperature (below 100 degrees C) and decarbonizing at about 300 degrees C-400 degrees C, the spongy ceramic block was sintered at high temperature, thus, macroporous HAp bioceramic with interconnected pores and reasonable porosity and pore-diameter was manufactured. This kind of porous HAp bioceramics were intrinsically osteoinductive to a certain degree, but its outstanding property was that they can absorb human bone morphogenetic proteins and other bone growth factors to form composites, so that the macroporous HAp bioactive ceramic has appropriate feasibility for clinical application. From the point of biomedical application, the recent developments in synthesis and characteristics investigation of macroporous HAp are reviewed in this paper.

5-Aminolevulinic acid esters based photodynamic therapy / 生物医学工程学杂志

As exogenous ALA (5-aminolevulinic acid) esters can induce the production and accumulation of endogenous photosensitizer PpIX (protoporphyrin IX) in tumor tissues more effectively, they have been the most active photosensitizer prodrug in PDT(photodynamic therapy) field. In this article, along with the procedure of ALA esters based PDT, some primary mechanism and experimental results were considered, which include: first, cellular uptake of ALA esters and its conversion into ALA; second, the production and accumulation of endogenous photosensitizer PpIX induced by eNdogenous ALA esters; last, the photosensitization of PpIX.