The Effect of Pulsed Direct Current Field on the Membrane Flux of a New Style of Membrane Bioreactor

The effect of a pulsed direct current electric field on the membrane flux of an antifouling Membrane Bioreactor (MBR) designed and built in the laboratory was studied. The results showed that a pulsed direct current electric field had a significant effect on the membrane flux. The average membrane flux when using a pulse direct current electric field was between that of a steady electric field and that of no electric current under a trans‐membrane pressure drop of ΔP = 0.1 MPa and a pulsed direct current electric field strength of 20 V/cm. There was an optimum value of pulse intensity and pulse duration for the membrane flux. The shorter the pulse interval time was, the higher the frequency of power‐on, and the higher the average stable membrane flux of the MBR. This may have resulted from the fact that charged coarse particles and colloids account for 80 % of COD_cr and BOD₅ in urban domestic sewage. These particles and colloids moved away from the membrane surface at a certain electrical field strength, gradually thinning the particle sedimentary layer, reducing the membrane filtration resistance, and increasing the membrane flux significantly. However, the formation of a sedimentary layer on the membrane surface needed a significant amount of time and an appropriate pulse frequency while the MBR is powered on. This could save energy while keeping the membrane clean.     

Effects of Pulsed Electromagnetic Field on Differentiation of HUES-17 Human Embryonic Stem Cell Line

Electromagnetic fields are considered to potentially affect embryonic development, but the mechanism is still unknown. In this study, human embryonic stem cell (hESC) line HUES-17 was applied to explore the mechanism of exposure on embryonic development to pulsed electromagnetic field (PEMF) for 400 pulses at different electric field intensities and the differentiation of HUES-17 cells was observed after PEMF exposure. The expression of alkaline phosphatase (AP), stage-specific embryonic antigen-3 (SSEA-3), SSEA-4 and the mRNA level and protein level of Oct4, Sox2 and Nanog in HUES-17 cells remained unchanged after PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m. Four hundred pulses PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m did not affect the differentiation of HUES-17 cells. The reason why electromagnetic fields affect embryonic development may be due to other mechanisms rather than affecting the differentiation of embryonic stem cells.     

人前列腺癌PC-3细胞膜蛋白组学的分析

目的分析人前列腺癌PC-3细胞中的所有膜蛋白,全面展示PC-3细胞中的蛋白质表达谱。方法采用一维SDS-PAGE,结合高效液相色谱-串联质谱的方法,分离并鉴定PC-3细胞中的膜蛋白。结果在严格的过滤参数条件下,PC-3细胞中鉴定出2172种蛋白,其中1499种经过基因本体评注(GOA)显示为已知细胞组分,其余为未知细胞组分。在已知细胞组分中,564种(37.6%)蛋白为质膜蛋白,其中138种为跨膜蛋白。跨膜蛋白中,21.17%被预测至少有1个跨膜区,57.66%有1～3个跨膜区,30.66%有4～9个跨膜区,11.66%有不少于10个跨膜区。许多新的蛋白也被鉴定,其中包括假设蛋白和一些cD-NA序列。结论这些被鉴定蛋白的生物学功能和理化性质将有助于进一步理解前列腺癌侵袭和转移的分子机制,为寻找与前列腺癌转移相关的分子提供新的实验证据。     

Effect of Pulsed Electric Field on Membrane Lipids and Oxidative Injury of Salmonella typhimurium

Salmonella typhimurium cells were subjected to pulsed electric field (PEF) treatment at 25 kV/cm for 0–4 ms to investigate the effect of PEF on the cytoplasmic membrane lipids and oxidative injury of cells. Results indicated that PEF treatment induced a decrease of membrane fluidity of Salmonella typhimurium (S. typhimuriumi), possibly due to the alterations of fatty acid biosynthesis-associated gene expressions (down-regulation of cfa and fabA gene expressions and the up-regulation of fabD gene expression), which, in turn, modified the composition of membrane lipid (decrease in the content ratio of unsaturated fatty acids to saturated fatty acids). In addition, oxidative injury induced by PEF treatment was associated with an increase in the content of malondialdehyde. The up-regulation of cytochrome bo oxidase gene expressions (cyoA, cyoB, and cyoC) indicated that membrane damage was induced by PEF treatment, which was related to the repairing mechanism of alleviating the oxidative injury caused by PEF treatment. Based on these results, we achieved better understanding of microbial injury induced by PEF, suggesting that micro-organisms tend to decrease membrane fluidity in response to PEF treatment and, thus, a greater membrane fluidity might improve the efficiency of PEF treatment to inactivate micro-organisms.     

Tumour Cell Membrane Poration and Ablation by Pulsed Low-Intensity Electric Field with Carbon Nanotubes

Electroporation is a physical method to increase permeabilization of cell membrane by electrical pulses. Carbon nanotubes (CNTs) can potentially act like “lighting rods” or exhibit direct physical force on cell membrane under alternating electromagnetic fields thus reducing the required field strength. A cell poration/ablation system was built for exploring these effects of CNTs in which two-electrode sets were constructed and two perpendicular electric fields could be generated sequentially. By applying this system to breast cancer cells in the presence of multi-walled CNTs (MWCNTs), the effective pulse amplitude was reduced to 50 V/cm (main field)/15 V/cm (alignment field) at the optimized pulse frequency (5 Hz) of 500 pulses. Under these conditions instant cell membrane permeabilization was increased to 38.62%, 2.77-fold higher than that without CNTs. Moreover, we also observed irreversible electroporation occurred under these conditions, such that only 39.23% of the cells were viable 24 h post treatment, in contrast to 87.01% cell viability without presence of CNTs. These results indicate that CNT-enhanced electroporation has the potential for tumour cell ablation by significantly lower electric fields than that in conventional electroporation therapy thus avoiding potential risks associated with the use of high intensity electric pulses.     

利用电泳迁移运动抑制膜面微粒沉积的研究

本文介绍了附加直流电场的十字流膜过滤，并着重探讨了附加电场使料浆中悬浮微粒产生电泳迁移的作用，作者还介绍了间断加电的研究，根据试验结果说明间断加电可望获得较好的节能效果。     

电场促进膜过滤技术研究进展

分离过程中常用的膜过滤多以错流方式进行,但是滤面的附着物和浓度极化会使过滤速度大大降低,研究表明电场能够促进过滤的进行。简要介绍了电场促进过滤的概念及原理,综述了国内外对含胶体或蛋白质等难溶物质的介质进行电场促进膜过滤方面所进行的理论和实验研究,重点是外加电场的方式、大小、脉冲电压的频率等因素对过滤速度的影响。     

Testing pulsed electric fields in electroremediation of copper mine tailings

Mining processes, prior to pyrometallurgical copper production, generate large amounts of solid waste, which are and have been deposited in mine tailings impoundments. This work is based on three electrodialytic (EDR) and four electrokinetic (EKR) remediation experiments on fresh and aged copper mine tailings using pulsed electric fields. The EDR results are compared to conventional remediation with direct current (dc) electric field. Applying pulsed electric fields in EDR, it was found that the remediation time decreased compared to dc EDR. In the anode side of the cell, 72 h of remediation at 20 V with a frequency ≥28 cycles/day at a fixed time ratio (t_ON/t_OFF) of 20, in terms of copper removal would correspond to around 270 h of dc remediation time. With both EDR and EKR of 120 h, at 14 cycles/day, the copper removal is practically uniform in the whole cell. Increasing the frequency to ≥28 cycles/day, the copper removal in the anode side is improved, and in the cathode side an accumulation is observed. With an EKR of 240 h, a larger copper accumulation in the cathode side was observed due to precipitation of copper hydroxide. In EDR with a pulsed electric field, the transport across the cation exchange membrane becomes the rate determining step. This can be seen from: (a) accumulation in the cathode side when increasing the frequency and (b) similar total copper removal for the three frequencies studied. In EKR, free alkali diffusion into the cell occurred. This generated copper precipitation in the cathode side, and the effect of the pulsed electric field on total copper removal was reduced.     

Initiation of Ignition by the Action of a High‐Current Pulsed Discharge on a Gas

The possibility of nonthermal initiation of chemical reactions by a uniform pulsed nanosecond discharge is demonstrated. Dependences of variation of the ignition delay on initial conditions are obtained. It is shown that the main role in combustion initiation under conditions of a pulsed gas discharge in the case of moderate electric fields and low degree of ionization belongs to reactions of dissociation quenching of electronexcited levels of nitrogen.     

Helicobacter pylori Disrupts Host Cell Membranes, Initiating a Repair Response and Cell Proliferation

Helicobacter pylori (H. pylori), the human stomach pathogen, lives on the inner surface of the stomach and causes chronic gastritis, peptic ulcer, and gastric cancer. Plasma membrane repair response is a matter of life and death for human cells against physical and biological damage. We here test the hypothesis that H. pylori also causes plasma membrane disruption injury, and that not only a membrane repair response but also a cell proliferation response are thereby activated. Vacuolating cytotoxin A (VacA) and cytotoxin-associated gene A (CagA) have been considered to be major H. pylori virulence factors. Gastric cancer cells were infected with H. pylori wild type (vacA+/cagA+), single mutant (ΔvacA or ΔcagA) or double mutant (ΔvacA/ΔcagA) strains and plasma membrane disruption events and consequent activation of membrane repair components monitored. H. pylori disrupts the host cell plasma membrane, allowing localized dye and extracellular Ca(2+) influx. Ca(2+)-triggered members of the annexin family, A1 and A4, translocate, in response to injury, to the plasma membrane, and cell surface expression of an exocytotic maker of repair, LAMP-2, increases. Additional forms of plasma membrane disruption, unrelated to H. pylori exposure, also promote host cell proliferation. We propose that H. pylori activation of a plasma membrane repair is pro-proliferative. This study might therefore provide new insight into potential mechanisms of H. pylori-induced gastric carcinogenesis.     

5-Fu对人结肠癌细胞凋亡过程中DNA损伤修复基因表达的影响

目的探讨5-Fu对人结肠癌细胞(HCT/8)凋亡过程中DNA损伤修复相关基因表达的影响。方法通过形态学、基因组电泳、吖啶橙染色及流式细胞仪,观察大肠癌细胞凋亡情况,同时应用基因芯片分析5-Fu作用后DNA损伤修复相关基因表达的变化。结果经流式细胞仪检测发现,5-Fu作用于HCT/8细胞48h与对照组相比,凋亡细胞数明显增加,基因组电泳出现典型的梯形条带,基因芯片分析显示,12个与DNA损伤修复相关的基因中,上调基因3个,下调基因9个。结论5-Fu作用于结肠癌细胞,导致多种DNA损伤修复基因发生改变,综合作用使细胞不能及时对损伤DNA进行修复,最终导致细胞发生凋亡。     

多孔PZT压电陶瓷膜的制备及其抗污染性能

以 PbZr_xTi_1-xO₃（PZT）压电陶瓷粉体为原料,通过干压成型的方法制备多孔PZT陶瓷膜,考察了煅烧温度对多孔PZT陶瓷膜的机械强度、孔隙率以及纯水渗透性能的影响。当煅烧温度为950℃时,可制备出纯水渗透率为850 L·m^-2·h^-1·MPa^-1,孔径为300 nm,机械强度为47.8 MPa,孔隙率为34%的多孔PZT陶瓷膜。在此基础上,考察了极化温度与极化电压对多孔PZT陶瓷膜压电性能的影响,并对极化后的PZT压电陶瓷膜进行萃取和表面等离子刻蚀处理。结果表明：极化温度为120℃、极化电压强度为4 kV·mm^-1,极化后经热乙醇萃取及表面等离子刻蚀4 min后,多孔PZT压电陶瓷膜在外加交流电为20 V时,产生的共振振幅信号值达34.8 mV。将制备的多孔PZT压电陶瓷膜在粒径为600 nm的含油乳化液中进行过滤实验,发现陶瓷膜两端未加交流电时,其通量在2 h内衰减至4%。而加交流电后,其稳定通量可维持在20%左右,表明制备的多孔PZT压电陶瓷膜具有良好的抗污染效果。     

电场强化错流膜过滤的研究

电场强化错流膜过滤技术可有效改善膜污染和浓差极化对错流过滤带来的不利影响。就电场膜过滤装置、机理、外加电场及影响渗透通量的因素这4个方面进行综述。目前研制的新型附加电场中空纤维膜组件克服了传统膜过滤组件的缺点,显示出很大的工程应用前景。电场强化错流膜过滤中会发生电泳和电渗等电动力学效应以及电化学效应。电泳是减缓膜污染一个主要因素,电泳和电渗两者共同作用,可以大幅提高过滤通量。采用脉冲电场可以减缓过滤速率的衰减幅度,节省单位体积滤液的电功率比耗。当颗粒具有较高Zeta电位、外加场强及跨膜压力接近临界值、错流速率较高时,过滤通量极限值将会得到提高。     

DNA甲基化对调控胰岛分化基因表达的影响

目的探讨DNA甲基化对调控胰岛分化基因表达的影响,为调控干细胞分化为胰岛细胞提供理论依据。方法采用甲基化DNA免疫共沉淀-实时定量PCR法(MeDIP-qPCR)检测129/J小鼠胚胎干细胞、NIT1细胞及NIH3T3细胞中Pdx-1、MafA、Nkx6.1和Oct4四种调控胰岛分化基因的DNA甲基化程度;同时采用实时定量RT-PCR检测上述3种细胞中4种基因mRNA表达水平,分析这些基因DNA甲基化水平差异与基因表达之间的关系。结果 Pdx-1、MafA和Nkx6.1基因在129/J小鼠胚胎干细胞和NIT1细胞中呈低甲基化,在NIH3T3细胞中则呈高甲基化,前两种细胞的甲基化程度明显低于后者(P<0.05);Oct4基因在129/J小鼠胚胎干细胞中未甲基化,在NIT1和NIH3T3细胞中呈低甲基化,NIT1细胞的甲基化程度明显低于NIH3T3细胞(P<0.05)。低甲基化的Pdx-1、MafA和Nkx6.1基因在NIT1细胞中可高效表达,而在NIH3T3和129/J小鼠胚胎干细胞中则未见表达(P<0.05);Oct4基因在129/J小鼠胚胎干细胞中可高效表达,在NIT1和NIH3T3细胞中则未见表达。结论转录起始区DNA甲基化程度可影响Pdx-1、MafA、Nkx6.1和Oct4基因的表达,参与β细胞的分化过程。     

Pulse-enhanced electrokinetic remediation of fluorine-contaminated soil

The presence of abundant fluorides in soil can pose potential threats to environment and human health. We investigated the effects of pulsed electric field on electrokinetic remediation of fluorine-contaminated soil by varying the pulse interval. Although the remediation time was increased, experimental results indicated pulse-enhanced electrokinetic remediation could improve the removal efficiency of fluorine better than the conventional electrokinetic remediation in the same intensity of electric field. The fluorine removal efficiency of the former (30 min, pulse interval) was 80.93% and that of normal electrokinetic remediation was 72.40%, because pulsed electric field could reduce the concentration polarization and increase the electrical current, electroosmotic flow and the electrical voltage part in soil cell. Therefore, in the later phase of electrokinetic remediation, when electrical current and the fluorine removal efficiency decreased obviously, at this time the pulse-enhanced method would be a considerable choice to eliminate fluorine in contaminated soils.     

电场强化陶瓷膜超滤牛血清蛋白

以相对分子质量67 000的牛血清蛋白(BSA)为分离对象,通过膜通量、截留率、膜通量衰减程度和恢复率等膜参数的测定,考察了外加直流电场作用陶瓷膜(γ-Al2O3)的分离性能。结果表明,外加正电场陶瓷膜的膜通量和截留率都有所提高,20 V/cm时,分别提高了67%和46.8%,而膜通量衰减程度由84.64%降低到60.87%,经过"电场自净"的陶瓷膜,其膜通量的恢复率可达95%以上,能较大改善膜的抗污染性能。     

Genes Responsive to Low-Intensity Pulsed Ultrasound in MC3T3-E1 Preosteoblast Cells

Although low-intensity pulsed ultrasound (LIPUS) has been shown to enhance bone fracture healing, the underlying mechanism of LIPUS remains to be fully elucidated. Here, to better understand the molecular mechanism underlying cellular responses to LIPUS, we investigated gene expression profiles in mouse MC3T3-E1 preosteoblast cells exposed to LIPUS using high-density oligonucleotide microarrays and computational gene expression analysis tools. Although treatment of the cells with a single 20-min LIPUS (1.5 MHz, 30 mW/cm²) did not affect the cell growth or alkaline phosphatase activity, the treatment significantly increased the mRNA level of Bglap. Microarray analysis demonstrated that 38 genes were upregulated and 37 genes were downregulated by 1.5-fold or more in the cells at 24-h post-treatment. Ingenuity pathway analysis demonstrated that the gene network U (up) contained many upregulated genes that were mainly associated with bone morphology in the category of biological functions of skeletal and muscular system development and function. Moreover, the biological function of the gene network D (down), which contained downregulated genes, was associated with gene expression, the cell cycle and connective tissue development and function. These results should help to further clarify the molecular basis of the mechanisms of the LIPUS response in osteoblast cells.     

Synergistic Effect of PVDF-Coated PCL-TCP Scaffolds and Pulsed Electromagnetic Field on Osteogenesis

Bone exhibits piezoelectric properties. Thus, electrical stimulations such as pulsed electromagnetic fields (PEMFs) and stimuli-responsive piezoelectric properties of scaffolds have been investigated separately to evaluate their efficacy in supporting osteogenesis. However, current understanding of cells responding under the combined influence of PEMF and piezoelectric properties in scaffolds is still lacking. Therefore, in this study, we fabricated piezoelectric scaffolds by functionalization of polycaprolactone-tricalcium phosphate (PCL-TCP) films with a polyvinylidene fluoride (PVDF) coating that is self-polarized by a modified breath-figure technique. The osteoinductive properties of these PVDF-coated PCL-TCP films on MC3T3-E1 cells were studied under the stimulation of PEMF. Piezoelectric and ferroelectric characterization demonstrated that scaffolds with piezoelectric coefficient d₃₃ = −1.2 pC/N were obtained at a powder dissolution temperature of 100 °C and coating relative humidity (RH) of 56%. DNA quantification showed that cell proliferation was significantly enhanced by PEMF as low as 0.6 mT and 50 Hz. Hydroxyapatite staining showed that cell mineralization was significantly enhanced by incorporation of PVDF coating. Gene expression study showed that the combination of PEMF and PVDF coating promoted late osteogenic gene expression marker most significantly. Collectively, our results suggest that the synergistic effects of PEMF and piezoelectric scaffolds on osteogenesis provide a promising alternative strategy for electrically augmented osteoinduction. The piezoelectric response of PVDF by PEMF, which could provide mechanical strain, is particularly interesting as it could deliver local mechanical stimulation to osteogenic cells using PEMF.