Lattice Boltzmann simulation study of anode degradation in solid oxide fuel cells during the initial aging process

For present solid oxide fuel cells(SOFCs),rapid performance degradation is observed in the initial aging process,and the dis-cussion of the degradation mechanism necessitates quantitative analysis.Herein,focused ion beam-scanning electron microscopy was em-ployed to characterize and reconstruct the ceramic microstructures of SOFC anodes.The lattice Boltzmann method(LBM)simulation of multiphysical and electrochemical processes in the reconstructed models was performed.Two samples collected from industrial-size cells were characterized,including a reduced reference cell and a cell with an initial aging process.Statistical parameters of the reconstructed microstructures revealed a significant decrease in the active triple-phase boundary and Ni connectivity in the aged cell compared with the reference cell.The LBM simulation revealed that activity degradation is dominant compared with microstructural degradation during the initial aging process,and the electrochemical reactions spread to the support layer in the aged cell.The microstructural and activity de-gradations are attributed to Ni migration and coarsening.

TiO₂Nanoparticles Induced Genotoxicity in Cultured Cells Using Atmospheric Scanning Electron Microscopy (ASEM)

Nano-sized titanium oxide nanoparticles (TiO2 NPs) are widely used as a dye in food and cosmetics. TiO2 NPs are known to induce DNA damage when incorporated into cells. However, no bioassay is currently available to easily determine the cell incorporation of TiO2 NPs or related DNA damage, and to date, few studies have examined the different degrees of incorporation into cells according to the size of the TiO2 NPs particles and the presence or absence of cell specificity regarding DNA damage. This present study was therefore designed to examine COS7 cells that had incorporated TiO2 NPs using atmospheric scanning electron microscopy (ASEM). The results indicated that absorption of TiO2 NPs into cells and nuclear abnormalities had occurred. ASEM is a rapid and simple technique that enables the observation of samples immediately after fixation with glutaraldehyde and staining with phosphotungstic acid, and this method was suggested to be useful in screening for DNA damage.

Electron microscopic observations and DNA chain fragmentation studies on apoptosis in bone tumor cells induced by ^(153)Sm-EDTMP

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Recent progress on advanced transmission electron microscopy characterization for halide perovskite semiconductors

Halide perovskites are strategically important in the field of energy materials. Along with the rapid development of the materials and related devices, there is an urgent need to understand the structure–property relationship from nanoscale to atomic scale. Much effort has been made in the past few years to overcome the difficulty of imaging limited by electron dose,and to further extend the investigation towards operando conditions. This review is dedicated to recent studies of advanced transmission electron microscopy(TEM) characterizations for halide perovskites. The irradiation damage caused by the interaction of electron beams and perovskites under conventional imaging conditions are first summarized and discussed. Low-dose TEM is then discussed, including electron diffraction and emerging techniques for high-resolution TEM(HRTEM) imaging. Atomic-resolution imaging, defects identification and chemical mapping on halide perovskites are reviewed. Cryo-TEM for halide perovskites is discussed, since it can readily suppress irradiation damage and has been rapidly developed in the past few years. Finally, the applications of in-situ TEM in the degradation study of perovskites under environmental conditions such as heating,biasing, light illumination and humidity are reviewed. More applications of emerging TEM characterizations are foreseen in the coming future, unveiling the structural origin of halide perovskite’s unique properties and degradation mechanism under operando conditions, so to assist the design of a more efficient and robust energy material.

Correlative microscopy of Purkinje cells

The Purkinje cell and their synaptic contacts have been described using(1)light microsocopy,(2)transmission and scanning electron microscopy,and freeze etching technique,(3)conventional and field emission scanning electron microscopy and cryofracture methods,(4)confocal laser scanning microscopy using intravital stain FM64,and(5)immunocytochemical techniques for Synapsin-I,PSD9-5,GluR1 subunit of AMPA receptors,N-cadherin,and CamKII alpha.The outer surface and inner content of plasma membrane,cell organelles,cytoskeleton,nucleus,dendritic and axonal processes have been exposed and analyzed in a three-dimensional view.The intramembrane morphology,in bi-and three-dimensional views,and immunocytochemical labeling of synaptic contacts with parallel and climbing fibers,basket and stellate cell axons have been characterized.Freeze etching technique,field emission scanning microscopy and cryofracture methods,and GluR1 immunohistochemistry showed the morphology and localization of postsynaptic receptors.Purkinje cell shows N-cadherin and CamKII alpha immunoreactivity.The correlative microscopy approach provides a deeper understanding of structure and function of the Purkinje cell,a new three-dimensional outer and inner vision,a more detailed study of afferent and intrinsic synaptic junctions,and of intracortical circuits.

玉米苞叶组织细胞结构生物质碳的制备及显微结构的研究

采用一步热解法,使用常见的玉米果穗苞叶为原料,制备了保留玉米苞叶组织细胞结构的生物质碳膜,采用扫描电镜,透射电镜等技术手段对样品的形貌和结构进行了表征,样品表面有明显的维管束以及大量的气孔的存在。不添加任何导电剂和添加剂,热解碳膜直接作为工作电极使用,在100 mA·g^(-1)的电流密度下,200次循环后仍能获得218 mAh·g^(-1)的可逆比容量。本工作在生物质废弃物利用,以及储能材料开发等方面有参考价值。

斑马鱼视网膜微细结构及发育特征

目的探讨斑马鱼眼球和视网膜的微细结构及不同发育阶段的形态特征,为其作为视觉研究模型奠定基础。方法分别选取不同周龄的8组斑马鱼,每组各6条,共48条。利用光学显微镜和透射电子显微镜观察不同发育阶段的斑马鱼眼球结构,并测量视网膜各层厚度,分析其时空发育模式。从显微结构及超微结构上观察视网膜中各种细胞形态特征及神经连接方式,尤其是视杆细胞和视锥细胞的结构差异。结果斑马鱼视网膜可分为色素上皮层、视杆视锥层、外界膜、外核层、外网层、内核层、内网层、神经节细胞层、神经纤维层和内界膜等10层。与视锥细胞相比,视杆细胞的细胞核更小且电子密度更高。感光细胞终足整齐地排列在外网层,与水平细胞和双极细胞形成神经连接,并在其内观察到多条突触带。在斑马鱼视网膜中,最早发育的是神经节细胞层和内核层,随着斑马鱼的生长发育,视杆视锥层和色素上皮层厚度逐渐增长,10周左右视网膜的结构基本发育完成。结论斑马鱼视网膜形态结构典型,分层明显,神经细胞高度分化,外网层神经连接丰富。斑马鱼眼球发育特征与大多数哺乳动物相似。

原位冷冻电镜技术和可视蛋白质组学前沿

近年来,随着原位冷冻电镜技术和人工智能技术的不断发展,结构生物学的研究模式发生了重大转变。结构解析不再局限于分离提纯的单一生物大分子,而是在细胞甚至组织内原位直接解析生物大分子的高分辨率结构,并对亚细胞区域整体分子景观进行结构探索,从而更深刻地理解细胞与组织内生命活动的分子机制。通过展示细胞内不同蛋白质复合物的精细原位结构,可以进一步生成蛋白质组的可视化定量空间分子结构功能图谱,即可视蛋白质组学。以原位冷冻电镜技术为代表的新技术能够在空间水平阐明细胞内蛋白质组的三维相互作用网络,促进从整体上系统性地理解细胞内生物大分子结构功能与互作机制。为推动中国原位冷冻电镜技术和可视蛋白质组学的发展,本文总结了其研究模式和最新前沿进展,结合具体实例展示了新概念与新技术的先进性,并对发展前景进行了展望。

Observation on Apoptosis of Erythroid Cells Induced by Dexamethasone

Using splenic cells of BALB/c mice infected with Friend Leukemia Virus (FVA) as a model of erythroid cells, we investigated the action of dexamethasone (Dex) to induce apoptosis of the cells in the presence of erythropoietin (EPO)——an apoptotic inhibitor in developing erythroid cells. The result indicated that after treatment with a certain range of Dex concentrations and prolonged incubation, the cells were characterized by the occurrence of DNA ladders which appeared on agarose electrophoresis. Transmission electron microscopy showed that karyopyknosis, chromatin condensation, dilatation of the perinuclear space, karyorrhexis, cytoplasmic vacuolization and cell fragmentation appeared in the cells depending on the dose of Dex and the treatment time. These results suggest that Dex could induce apoptosis in the developing erythroid cells as in other cells so far reported.

Effects of tachyplesin on the morphology and ultrastructure of human gastric carcinoma cell line BGC-823

AIM To investigate the morphological andultrastructural changes in the human gastriccarcinoma cell line BGC-823 after being treatedwith tachyplesin.METHODS Tachyplesin was isolated from acidextracts of Chinese horseshoe crab(Tachypleustridentatus)hemocytes.BGC-823 cells and thecells treated with 2.0mg/L tachyplesin wereexamined respectively under light microscope,scanning and transmission electron microscope.RESULTS BGC-823 cells had undergone therestorational alteration in morphology andultrastructure after tachyplesin treatment.Thechanges were as follows:the shape of cells wasunanimous,the volume enlarged and cellsturned to be flat and spread,the nucleo-cytoplasmic ratio lessened and nuclear shapebecame rather regular,the number of nucleolusreduced and its volume lessened,heter-chromatin decreased while euchromatinincreased in nucleus.In the cytoplasm,mitochondria grew in number with consistentstructure relatively,Golgi complex turned to betypical and well-developed,rough endoplasmicreticulum increased and polyribosomedecreased.The microvilli at cellular surfacewere rare and the filopodia reduced whilelamellipodia increased at the cell edge.CONCLUSION Tachyplesin could alter themalignant morphological and ultrastructuralcharacteristics of human gastric carcinoma cellseffectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.

ETM study of electroporation influence on cell morphology in human malignant melanoma and human primary gingival fibroblast cells

Objective:To estimate electroporation(EP) influence on malignant and normal cells.Methods: Two cell lines including human malignant melanoma(Me-43) and normal human gingival fibroblast(HCFs) were used.EP parameters were the following:230,1000,1 730,2 300 V/cm;30 μ s by 3 impulses for every case.The viability of cells after EP was estimated by MTT assay. The ullrastructural analysis was observed by transmission electron microscope(Zeiss EM 900). Results:In the current study we observed the intracellular effect following EP on Me-43 and HGF cells.At the conditions applied,we did not observe any significant damage of mitochondrial activity in both cell lines treated by EP.Conversely,we showed that EP in some conditions can stimulate cells to proliferation.Some changes induced by EP were only visible in electron microscopy.In fibroblast cells we observed significant changes in lower parameters of EP(230 and 1 000 V/cm).After applying higher electric field intensities(2 300 V/cm) we detected many vacuoles,myelin-like bodies and swallowed endoplasmic reticulum.In melanoma cells such strong pathological modifications after EP were not observed,in comparison with control cells. The ultrastructure of both treated cell lines was changed according to the applied parameters of EP.Conclusions:We can claim that EP conditions are cell line dependent.In terms of the intracellular morphology,human fibroblasts are more sensitive to electric field as compared with melanoma cells.Optimal conditions should be determined for each cell line.Summarizing our study,we can conclude that EP is not an invasive method for human normal and malignant cells. This technique can be safely applied in chemotherapy for delivering drugs into tumor cells.

Mesenchymal stem cells and collagen patches for anterior cruciate ligament repair

AIM: To investigate collagen patches seeded with mesenchymal stem cells(MSCs) and/or tenocytes(TCs) with regards to their suitability for anterior cruciate ligament(ACL) repair. METHODS: Dynamic intraligamentary stabilization utilizes a dynamic screw system to keep ACL remnants in place and promote biological healing, supplemented by collagen patches. How these scaffolds interact with cells and what type of benefit they provide has not yet been investigated in detail. Primary ACL-derived TCs and human bone marrow derived MSCs were seeded onto two different types of 3D collagen scaffolds, Chondro-Gide?(CG) and Novocart?(NC). Cells were seeded onto the scaffolds and cultured for 7 d either as a pure populations or as "premix" containing a 1:1 ratio of TCs to MSCs. Additionally, as controls, cells were seeded in monolayers and in co-cultures on both sides of porous high-density membrane inserts(0.4 μm). We analyzed the patches by real time polymerase chain reaction, glycosaminoglycan(GAG), DNA and hydroxyproline(HYP) content. To determine cell spreading and adherence in the scaffolds microscopic imaging techniques, i.e., confocal laser scanning microscopy(c LSM) and scanning electron microscopy(SEM), were applied.RESULTS: CLSM and SEM imaging analysis confirmed cell adherence onto scaffolds. The metabolic cell activity revealed that patches promote adherence and proliferation of cells. The most dramatic increase in absolute metabolic cell activity was measured for CG samples seeded with tenocytes or a 1:1 cell premix. Analysis of DNA content and c LSM imaging also indicated MSCs were not proliferating as nicely as tenocytes on CG. The HYP to GAG ratio significantly changed for the premix group, resulting from a slightly lower GAG content, demonstrating that the cells are modifying the underlying matrix. Real-time quantitativepolymerase chain reaction data indicated that MSCs showed a trend of differentiation towards a more tenogenic-like phenotype after 7 d.CONCLUSION: CG and NC are both cyto-compatible with primary MSCs and TCs; TCs seemed to perform better on these collagen patches than MSCs.

Blockage of IGF-1R signaling sensitizes urinary bladder cancer cells to mitomycin-mediated cytotoxicity

A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signaling plays a very important role in progression, invasion and metastasis of bladder cancer cells. In this study, we investigated whether IGF-1R was involved in the growth stimulating activity and drug resistance of bladder cancer cells. The results showed: The mRNAs of IGF-1, IGF-2 and IGF-1R were strongly expressed in serum-free cultured T24 cell line, whereas normal urothelial cells did not express these factors/receptors or only in trace levels; T24 cell responded far better to growth stimulation by IGF-1 than did normal urothelial cells; blockage of IGF1R by antisense oligodeoxynucleotide (ODN) significantly inhibited the growth of T24 cell and enhanced sensitivity and apoptosis of T24 cells to mitomycin (MMC). These results suggested that blockage of IGF-IR signaling might potentially contribute to the treatment of bladder cancer cells which are insensitive to chemotherapy.

Esophageal cell proliferation in gastroesophageal reflux disease: Clinical-morphological data before and after pantoprazole

AIM: To evaluate esophageal mucosal defense mechanisms at an epithelial level to establish if pantoprazole treatment can induce ultrastructural healing and improvement in the proliferation activity of the esophageal epithelium in gastroesophageal reflux disease (GERD). METHODS: This was a single-blinded study for pHmonitoring, and histological, ultrastructural and MIB1 immunostaining evaluation. Fifty eight patients with GERD were enrolled and underwent 24 h pH-monitoring and endoscopy. Patients were treated for 12 and 24 mo with pantoprazole. Esophageal specimens were taken for histological and ultrastructural evaluation, before and after the treatment. RESULTS: With transmission electron microscopy, all patients with GERD showed ultrastructural signs of damage with dilation of intercellular spaces (DIS). After 3 mo of therapy the mean DIS values showed asignificant reduction and the mean MIB1-LI values of GERD showed an increase in cell proliferation. A further 3 mo of therapy significantly increased cell proliferation only in the erosive esophagitis (ERD) group. CONCLUSION: Three months of pantoprazole therapy induced ultrastructural healing of mucosal damage in 89% and 93% of ERD and non-erosion patients, respectively. Moreover, long-term pantoprazole treatment may be helpful in increasing the capability for esophageal cell proliferation in GERD, particularly in ERD patients.

A study of gentamicin injury mechanisms using cultured mouse cochlear spiral ganglion cells

Objective To study gentamicin injury mechanisms using postnatal mouse cochlear spiral gangcells （SGC）. Methods SGCs were isolated using a combinatorial approach of enzymatic digestion and mechanical separation from P2 - 6 Kunming mouse cochleae. After 4 days, cultured SGCs were fixed with 4% paraformaldehyde at room temperature for immunocytochemical examination using the methods of S-P and the monoclonal antibody against mouse neurofilament protein （Neurofilament-68/200Kda, NF-L＋ H）. SGCs were randomly divided into a blank control group and three gentamicin treatment groups （medium gentamicin concentration at 50 mg/L, 100 mg/L and 150 mg/L respectively）, SGCs were collected and examined under a transmission electron microscope after being cultured for 48 h. Results SGC primary culture was successful. SGC cytoplasm and neurites were dyed brownish yellow by the monoelonal mouse neurofilament protein antibody. SGCs showed classical bipolar neuron appearance. Under the transmission electron microscope,.gentamicin treated SGCs showed morphological features different compared to those in the blank control group, which might indicate apoptosis. Conclusion Our results indicate that gentamicin has direct toxic effects on cochlear SGCs in mice and the injury mechanism is closely related with apoptosis. Damage to mitochor, dria may play an important role in the process.

Influence of PTFE on Electrode Structure for Performance of PEMFC and 10-Cells Stack

Water plays a critical role on the performance, stability and lifetime of proton exchange membrane fuel cells（PEMFCs）. The addition of poly tetrafluoroethylene（PTFE） to the gas diffusion layer, especially, the cathode side, would optimize the transportation of water, electron and gas and thus improve the performance of the fuel cell. But until now, the studies about directly applying the PTFE to the catalyst layer are rarely reported. In this paper, the membrane electrode is fabricated by using directly coating catalyst to the membrane method（CCM） and applying PTFE directly to the cathode electrode catalyst layer. The performance of the single cell is determined by polarization curves and durability tests. Electrochemical impedance spectroscopy（EIS） and scanning electron microscopy（SEM） techniques are used to characterize the electrochemical properties of PEMFC. Also the performance of a 10-cells stack is detected. Combining the performance and the physical-chemistry characterization of PEMFC shows that addition of appropriate content of PTFE to the electrode enhances the performance of the fuel cell, which may be due to the improved water management. Addition of appropriate content of PTFE enhances the interaction between the membrane and the catalyst layer, and bigger pores and highly textured structure form in the MEA, which favors the oxygen mass transfer and protons transfer in the fuel cell. While superfluous addition of PTFE covers the surface of catalysts and hindered the contact of catalyst with Nation, which leads to the reduction of electrochemical active area and the decay of the fuel cell performance. The proposed research would optimize the water management of the fuel cell and thus improve the performance of the fuel cell.

Retrovirus-mediated herpes simplex virus thymidine kinase gene therapy approach for hepatocellular carcinoma

The therapeutic effect of herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) system on hepatocellular carcinoma was studied in this experiment. The tk-containing retroviral recombinants were used to infect hepatoma cells (BEL-7402) and the cells were treated with ganciclovir (0-1000 microg/ml). The results showed that HSV-tk gene could be efficiently transferred in vitro into hepatoma cells and stably expressed. The growth potential of the tk-containing cells was significantly inhibited by GCV (P

Influencing factors of rat small intestinal epithelial cell cultivation and effects of radiation on cell proliferation

INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisolated and cultured crypt cells from embryonalintestines and aseptic animal intestine, but failed.We introduced normal rat epithelial cell line-IEC-6from the USA and its living condition for stablepassage was successfully established after trials. Thecell line was testified to be the small intestinalepithelial cell by electron microscopy,immunihistochemistry and enzymatic histoch-emistry. It has been applied to some relatedresearch work[17-21]. It was found that manyfactors were involved in the culture system. Ourpresent study focuses on the culture method and theinfluencing factors on IEC-6.

Characteristics of an Established Retinoblastoma Cell Line HXO-Rb_(44)

Purpose:To study the retinoblastoma cell culture and to establish a new retinoblastoma cellline.Methods:22 retinoblastomes were cultured by using the method of single cellsus-pension.Characteristics of the cultured cells were studied in the following pro-grams:tumor cell morphology in vitro,electron microscopic,growth curve,cloning in soft agar,immunohistochemistry,karyotype and tumorigenicity.Results.22 retinoblastoas were cultured successfully in ivtro,only a cotinued cell line HXO-Rb44was established(more than3years).The characteristics of this cell line are that it grew as a suspension of round cell in graps like clusters in vitro,its population doubling time was 44hours,and it could be cloned in softa-gar.Histopathologic and ulatastructured pictures showed the characteristics of Rb.HXO-Rb44cell was positive to NSE and negative to GFAP in immunohis-tochemical staining.A subcutaneous injection of HXO-Rb44cells produced a retinoblastoma in BALB/C athumic nude mice.Conclusions:HXO-Rb44 has the characteristice of retinoblastoma and is a new retinoblastoma cell line.It is a useflu material for study this tumor both in basic and clinical fields.

Comparative ultrastructural study of endoplasmic reticulum in colorectal carcinoma cell lines with different degrees of differentiation

The endoplasmic reticulum (ER) consists of a complex system of tubules, lamellae, and flattened vesicles, and has a variety of morphologies in different cells. It is believed to play a central role in the biosynthesis of cholesterol, phospholipids, steroids, prostaglandins, membrane and secretory proteins[1]. Cancer cells have different functions and ultrastmcture from their original cells[2-4]. The studies on ER membrane system of cancer cells are of great significance in understanding their malignant behavior. In the present work, the ultrastructural characteristics of ER in human colorectal carcinoma cell lines with different differentiation degrees were investigated.