慢性家族性良性天疱疮的共聚激光扫描显微镜的影像特征

目的分析慢性家族性良性天疱疮患者皮损在共聚激光扫描显微镜(CLSM)下的影像特征。方法对入选患者的选定皮损处进行共聚激光扫描显微镜扫描,再取该处皮损进行组织病理检查,对这两者进行比较。结果皮损处的CLSM影像可见表皮棘层多处不规则断开或未完全断开,形成较多裂隙,棘层广泛松解,棘细胞间桥消失,但少数细胞间桥仍存在;因棘层松解而形成的腔隙中有单个或成团脱落的棘层松解细胞,基底细胞层有"绒毛"形成。结论慢性家族性良性天疱疮患者CLSM影像具有特征性表现,但该研究样本数有限,还需进一步研究论证。

Solidification process of conventional superalloy by confocal scanning laser microscope

The solidification process of a conventional superalloy, IN718, was investigated by confocal scanning laser microscope （CSLM）. The liquid fraction during solidification was obtained as a function of real time and temperature in reference with the in-situ observation. The characteristics of L→γ transformation were analyzed and the γ growing rate of each stage was also calculated. Scheil equation was employed to predict the segregation behavior, and the predict results are in consistence with the experimental results. As a result, the confocal scanning laser microscope shows a great potential for solidification process research.

胃癌的自体荧光图像分析

探讨胃癌组织显微自体荧光图像的特征表现。方法：采用氖离子（Ａｒ＋）激光（激发波长 ４８８ｎｍ）和氦氖(Ｈｅ－Ｎｅ）激光（激发波长５４３ ｎｍ）的双通道法激光扫描共聚焦显微镜对１６例胃癌手术标本进行自体荧光图像分析，井作自身对照研究。结果：胃正常组织各层的显微自体荧光图像均以绿色荧光为主。胃癌的显微自体荧光图像则红色荧光强度明显增加，并可见棕红色区，其显示率达１００％。结论：棕红色区是胃癌在显微自体荧光图像上的特征表现，自体荧光图像分析是胃癌诊断的有效方法。

Revealing the F_actin Networks in Interphase Nuclei of Garlic Clove Cells by Confocal Fluorescence Microscopy

The interphase nuclei of parenchyma cells and epidermal cells of garlic ( Allium sativum L.) clove were labelled with rabbit anti_actin antibody and FITC_conjugated goat anti_rabbit IgG antibody. The authors observed results with fluorescence microscopy and confocal laser scanning microscopy. The nuclei showed prominent green_yellow fluorescence, indicating the presence of actin in the nuclei. Fluorescence examination with TRITC_phalloidin showed distinctive red fluorescence in the nuclei, indicating that F_actin is present in the nuclei. Confocal laser scanning microscopy indicated the presence of F_actin containing network structures in the nuclei, but the network structures were absent and the nuclei still showed red fluorescence when the cells were treated with cytochalasin D before fixation; the red fluorescence in the nuclei was hard to be observed when the cells were treated with unlabelled phalloidin before the cells were stained with TRITC_phalloidin. These results indicate that F_actin is in the nuclei and forms network structures in the nuclei of garlic cells.

Colonization Pattern of Azospirillum brasilense Yu62 on Maize Roots

Plasmid pVK1001 which carried the gfp gene of GFPmut2, a mutant of GFP, was introduced into Azospirillum brasilense Yu62 by electroporation. Maize seedlings were inoculated with the GFP-labelled baeteria and grown gnotobiotically in flask with semi-solid agar medium. Observations were performed with confocal laser scanning microscopy (CLSM) and electron microscopy, respectively, at 8 d and 12 d after inoculation. Confocal laser scanning microscopy showed that A. brasilense Yu62 could penetrate into the cortex tissue, colonizing in the intercellular spaces of the parenchyma cells of the cortex tissue. Transmission and scanning electron microscopy (TEM) showed that the majority of the bacteria colonized on the root surface and only a minority of them resided in the root interior.

絮凝过程工艺设计的一种新概念

本文着重研究了在不同条件下所产生的絮团的内部结构。选用了两种不同的颗粒浓度 ,以改变原生颗粒或微絮团形成过程中微絮团碰撞几率。用共焦激光扫描显微镜作为一种温和的非破坏性技术 ,对几种典型的聚集体进行了测定 ,以获得有关透明的颗粒絮凝体的内部结构信息。然后用三维图象分析软件工具对获得的信息进行处理和分析 ,最后再用专门改进过的群集分析算法进行处理。用统计图来解释和比较显著水平很高的两个试验中絮凝体碰撞的最后阶段。由较高浓度的悬浮液形成的大絮团中具有较多数目的微絮团。由此可见 ,用这种试验技术和数学方法有助于更深入地了解絮团成因 。

Changes of the Microtubule Arrays During Mitosis in Prothallus Cells of Dryopteris crassirhizoma

Microtubule arrays in prothalli large-vacuolated and meristematic dividing cells of the fern Dryopteris crassirhizoma Nakai were studied using Steedman's wax, indirect immunofluorescence labelling and confocal laser scanning microscopy. Results showed that the use of high paraformaldehyde concentration (8%) allowed good fixation of prothallus cells, which are characterized by numerous (meristematic cells) and big (large-vacuolated cells) vacuoles. Results also plead for the efficiency of Steedman's wax embedding method in: (1) avoiding excessive use of enzyme for digesting cell wall in the process of the microtubule cytoskeleton labelling, (2) minimizing the autofluorescence effect in cells through utilization of alcohol in sample dehydration, and (3) permitting a clear visualization of microtubule patterns during the cell mitosis. Steedman's wax, coupled with immunofluorescence labelling and confocal laser scanning microscopy techniques, allows a good investigation of cell division process in plants by using simple multicellular organisms such as fern prothalli.

Effect of current density on corrosion resistance of micro-arc oxide coatings on magnesium alloy

Oxide coatings were prepared on magnesium alloys in electrolyte solution of Na2SiO3 at different current densities(3,4 and 5 A/cm 2 )with micro-arc oxidation process.X-ray diffractometry(XRD)results show that the oxide coatings formed on magnesium alloys are mainly composed of MgO and MgAl2O4 phases;in addition,the content of MgO increases with increasing the current density.The morphology and surface roughness of the coatings were characterized by confocal laser scanning microscopy (CLSM).The results show that the surface roughness(Ra)decreases with increasing the current density.Moreover,the electrochemical corrosion results prove that the MgO coating produced in the electrolyte Na2SiO3 at current density of 5 A/cm 2 shows the best corrosion resistance.

Confocal laser endomicroscopy in the “in vivo” histological diagnosis of the gastrointestinal tract

Recent technological advances in miniaturization have allowed for a confocal scanning microscope to be integrated into a conventional flexible endoscope,or into trans-endoscopic probes,a technique now known as confocal endomicroscopy or confocal laser endomicroscopy.This newly-developed technology has enabled endoscopists to collect real-time in vivo histological images or "virtual biopsies" of the gastrointestinal mucosa during endoscopy,and has stimulated significant interest in the application of this technique in clinical gastroenterology.This review aims to evaluate the current data on the technical aspects and the utility of this new technology in clinical gastroenterology and its potential impact in the future,particularly in the screening or surveillance of gastrointestinal neoplasia.

A Confocal Endoscope for Cellular Imaging

Since its inception, endoscopy has aimed to establish an immediate diagnosis that is virtually consistent with a histologic diagnosis. In the past decade, confocal laser scanning microscopy has been brought into endoscopy, thus enabling in vivo microscopic tissue visualization with a magnification and resolution comparable to that obtained with the ex vivo microscopy of histological specimens. The major challenge in the development of instrumentation lies in the miniaturization of a fiber-optic probe for microscopic imaging with micron-scale resolution. Here, we present the design and construction of a confocal endoscope based on a fiber bundle with 1.4-μm lateral resolution and 8-frames per second(fps) imaging speed. The fiber-optic probe has a diameter of 2.6 mm that is compatible with the biopsy channel of a conventional endoscope. The prototype of a confocal endoscope has been used to observe epithelial cells of the gastrointestinal tracts of mice and will be further demonstrated in clinical trials. In addition, the confocal endoscope can be used for translational studies of epithelial function in order to monitor how molecules work and how cells interact in their natural environment.

Visualization of Golgia apparatus as an intracellular calcium store by laser scanning confocal microscope

Using laser scanning confocal microscopy, we have found that the in cells loaded with fluo-3/AM, highest intracellular Ca(2+) in the perinuclear region is associated with the Golgi apparatus. The spatiotemporal subcellu lar distribution of Ca(2+) in living human fibroblasts exposing to calcium-free medium in response to agonists has been investigated. PDGF, which releases Ca(2+) from intracellular stores by inositol(1, 4, 5)-trisphosphate pathway,produced a biphasic transient rise in intracellular calcium.The initial rise was resulted from a direct release of calcium from the Golgi apparatus. Calcium could be also released from and reaccumulated into the Golgi apparatus by the stimulation of thapsigargin, an inhibitor of the Ca(2+) transport ATPase of intracellular calcium store. Permeablizing the plasma membrane by 10 μM digitonin resulted in the calcium release from the Golgi apparatus and depletion of the internal calcium store. These results suggest that the Golgi apparatus plays a role in Ca(2+) regulation in signal transduction.

Choice of reference surfaces for machined surface roughness in milling of SiC_p/Al composites

In order to choose the appropriate reference surface on the machined surface roughness of Si Cp/Al composites, the cutting experiments of Si Cp/Al composites were carried out, and the machined surface topography was measured by OLS3000 Confocal laser scanning microscope. The 3D measured data of machined surface topography were analyzed by the area power spectrum density. The result shows that the texture of machined surface topography in milling of Si Cp/Al composites is almost isotropic. This is the reason that the values of Rq at different locations on the same machined surface are obviously different. Through the comparison of performance of different filtering methods, the robust least squares reference surface can be used to extract the surface roughness of SiC p/Al composites effectively.

Optimization of four types of antimicrobial agents to increase the inhibitory ability of marine Arthrobacter oxydans KQ 11 dextranase mouthwash

We adopted the response surface methodology using single factor and orthogonal experiments to optimize four types of antimicrobial agents that could inhibit biofilm formation by Streptococcus mutans, which is commonly found in the human oral cavity and causes tooth decay. The objective was to improve the function of marine Arthrobacter oxydans KQll dextranase mouthwash （designed and developed by our laboratory）. The experiment was conducted in a three-level, four-variable central composite design to determine the best combination of ZnSO4, lysozyme, citric acid and chitosan. The optimized antibacterial agents were 2.16 g/L ZnSO4, 14 g/L lysozyme, 4.5 g/L citric acid and 5 g/L chitosan. The biofilm formation inhibition reached 84.49%. In addition, microscopic observation of the biofilm was performed using scanning electron microscopy and confocal laser scanning microscopy. The optimized formula was tested in marine dextranase Arthrobacter oxydans KQ11 mouthwash and enhanced the inhibition of S. mutans. This work may be promoted for the design and development of future marine dextranase oral care products.

Effects of Corrosion Inhibitors on Lubrication Performance of Rolling Oil for Copper Foil

The 2,5-bis（ethyldisulfanyl）-l,3,4-thiadiazole （T561）, benzotriazole （BTA）,1-N, N-bis （2-ethylhexyl） aminomethyl-4-methyl-lh-benzotriazole （IRGAMET39） and I-IN, N-bis （2-ethylhexyl） aminomethyl] methyl benzotriazole （TT- LX） have been evaluated as corrosion inhibitors used in rolling oil for cold rolling of copper foil. The MRS-10A four-ball friction and wear tests have been carried out to compare their tribological properties, and the lubricating performance of rolling oils has been studied through rolling experiments. The oil sample containing IRGAMET 39 has the same PB value as that one containing T561, with the coefficient of friction increased by 35.6% and wear scar diameter decreased by 4%. The minimum rolling gauge has been studied after rolling lubrication, but the results show that inhibitors have no effect on it. Scanning electron microscopy （SEM） and energy dispersive spectrometry （EDS） analyses have indicated that the inhibitor is adsorbed on the copper surface to prevent copper from being corroded easily. In addition, the LEXT OLS4000 laser confocal microscopy has been used to observe the foil surface which shows that the streaks of foil surface are clear, the scratches are shallow and the surface failure is improved effectively.

Mechanisms of cholecystokinin-induced calcium mobilization in gastric antral interstitial cells of Cajal

AIM:To investigate the effect of sulfated cholecystokinin-8(CCK-8S) on calcium mobilization in cultured murine gastric antral interstitial cells of Cajal(ICC) and its possible mechanisms.METHODS:ICC were isolated from the gastric antrum of mice and cultured.Immunofluorescence staining with a monoclonal antibody for c-Kit was used to identify ICC.The responsiveness of ICC to CCK-8S was measured using Fluo-3/AM based digital microfluorimetric measurement of intracellular Ca2+ concentration([Ca2+]i).A confocal laser scanning microscope was used to monitor [Ca2+]i changes.The selective CCK1 receptor antagonist lorglumide,the intracellular Ca2+-ATPase inhibitor thapsigargin,the type Ⅲ inositol 1,4,5-triphosphate(InsP3) receptor blocker xestospongin C and the L-type voltage-operated Ca2+ channel inhibitor nifedipine were used to examine the mecha-nisms of [Ca2+]i elevation caused by CCK-8S.Immunoprecipitation and Western blotting were used to determine the regulatory effect of PKC on phosphorylation of type Ⅲ InsP3 receptor(InsP3R3) in ICC.Protein kinase C(PKC) activator phorbol 12-myristate 13-acetate(PMA) and inhibitor chelerythrine were used to assess the role of PKC in the CCK-8S-evoked [Ca2+]i increment of ICC.RESULTS:ICC were successfully isolated from the gastric antrum of mice and cultured.Cultured ICC were identified by immunofluorescence staining.When given 80 nmol/L or more than 80 nmol/L CCK-8S,the [Ca2+]i in ICC increased and 100 nmol/L CCK-8S significantly increased the mean [Ca2+]i by 59.30% ± 4.85%(P < 0.01).Pretreatment of ICC with 5 μmol/L lorglumide inhibited 100 nmol/L CCK-8S-induced [Ca2+]i increment from 59.30% ± 4.85% to 14.97% ± 9.05%(P < 0.01),suggesting a CCK1R-mediated event.Emptying of intracellular calcium stores by thapsigargin(5 μmol/L) prevented CCK-8S(100 nmol/L) from inducing a [Ca2+]i increase.Moreover,pretreatment with xestospongin C(1 μmol/L) could also abolish the CCK-8S-induced effect,indicating that Ca2+ release from InsP3R-operated stores appeared to be a major mechanism responsible for CCK-8S-induced calcium mobilization in ICC.On the other hand,by removing extracellular calcium or blocking the L-type voltage-operated calcium channel with nifedipine,a smaller but significant rise in the [Ca2+]i could be still elicited by CCK-8S.These data suggest that the [Ca2+]i release is not stimulated or activated by the influx of extracellular Ca2+ in ICC,but the influx of extracellular Ca2+ can facilitate the [Ca2+]i increase evoked by CCK-8S.CCK-8S increased the phosphorylation of InsP3R3,which could be prevented by chelerythrine.Pretreatment with lorglumide(5 μmol/L) could significantly reduce the CCK-8S intensified phosphorylation of InsP3R3.In the positive control group,treatment of cells with PMA also resulted in an enhanced phosphorylation of InsP3R3.Pretreatment with various concentrations of PMA(10 nmol/L-10 μmol/L) apparently inhibited the effect of CCK-8S and the effect of100 nmol/L PMA was most obvious.Likewise,the effect of CCK-8S was augmented by the pretreatment with chelerythrine(10 nmol/L-10 μmol/L) and 100 nmol/L chelerythrine exhibited the maximum effect.CONCLUSION:CCK-8S increases [Ca2+]i in ICC via the CCK1 receptor.This effect depends on the release of InsP3R-operated Ca2+ stores,which is negatively regulated by PKC-mediated phosphorylation of InsP3R3.

Observation on Double Fertilization and Early Embryonic Development in Autotetraploid Polyembryonic Rice

The process of double fertilization and the characters of embryo and endosperm development in an autotetraploid polyembryonic mutant rice IR36-Shuang were studied with a laser scanning confocal microscopy. Some abnormalities including degenerated ovary, abortive embryo sac, single fertilization, double-ovule and double-embryo and so on. were found dudng double fertilization and embryo development in IR36-Shuang. The rate of the abnormalities was 46.67% in IR36-Shuang, significantly higher than that in the control, an autotetraploid rice line IR36-4X （33.00%）. Cytological and embryonic evidences were provided for seed setting decline and the initiation of additional embryo in IR36-Shuang.

Cxcl16 Interact With SARS-CoV N Protein In and Out Cell

Our study investigated the host cell protein which can interact with SARS-CoV N protein, and explored the functional connections. The eukaryotic expression vectors pEGFP-N1/SARS-CoVN and pdsRed2-N1/ CXCL16 were constructed and used to co-transfect HEK293FT cells by the calcium phosphate method. The HIS-tagged fusion protein SARS-CoVN-GFP was then built and purified for the binding assay in vitro. The co-localization of SARS-CoVN and CXCL16 in the cytoplasm of HEK293FT cells was also shown using confocal laser scanning microscopy. It is suggested that their interaction might be through direct combination. Under a fluorescence microscope, it was observed that the purified fusion protein SARS-CoVN-GFP was attached to the cell membrane of CXCL16-transfected cells, indicating that SARS-CoVN and CXCL16 can be mutually combined.

Suppression of gold nanoparticle agglomeration and its separation via nylon membranes

Use of ultraporous nylon membrane is one of the most widely employed techniques for removal of hard and soft nanoparticles in the semiconductor industry, and the accurate determination of membrane pore size is necessary in order to avoid manufacturing defects caused by contamination. The gold nanoparticle has several benefits for the evaluation of polymeric membranes; however, the nanoparticles agglomerate easily on the nylon membrane and make it difficult to evaluate the membrane precisely. The properties of 2-amino-2-hydroxymethyl-1,3-propanediol(ADP) ligand in gold nanoparticle solution were systematically investigated, and ADP was utilized for improved evaluation of the nylon membranes. Nylon membrane used in this study was prepared by phase inversion techniques. Ultrathin dense layer on top of the membrane surface and Darcy structures in the microporous membrane support were observed. The gold particle rejection was carried out at various p H values from 4 to14 and higher rejection was observed at p H 4 and 8. The suppression of gold colloid agglomeration using ADP and monodispersity of gold colloids was also analyzed by confocal laser scanning microscopy(CLSM), transmission electron microscopy(TEM), and scanning electron microscopy(SEM). van der Waals interaction energy of the particles was reduced in the addition of ADP. The presence of ADP ligand in the gold solutions prevented the agglomeration of gold nanoparticles and reduced the adsorption of the particles on the nylon membrane surface,leading to precise evaluation of membrane pore sizes.

Surface Functionalization of Microporous Polypropylene Membrane with Polyols for Removal of Boron Acid from Aqueous Solution

Affinity membranes are fabricated for boric acid removal by the surface functionalization of microporous polypropylene membrane(MPPM)with lactose-based polyols.The affinity is based on specific complexation between boric acid and saccharide polyols.A photoinduced grafting-chemical reaction sequence was used to prepare these affinity membranes.Poly(2-aminoethyl methacrylate hydrochloride)[poly(AEMA)]was grafted on the surfaces of MPPM by UV-induced graft polymerization.Grafting in the membrane pores was visualized by dying the cross-section of poly(AEMA)-grafted MPPM with fluorescein disodium and imaging with confocal laser scanning microscopy.It is concluded that lactose ligands can be covalently immobilized on the external surface and in the pores by the subsequent coupling of poly(AEMA)with lactobionic acid(LA).Physical and chemical properties of the affinity membranes were characterized by field emission scanning electron microscopy and Fourier Transform Infrared/Attenuated Total Refraction spectroscopy(FT-IR/ATR).3-Aminophenyl boric acid(3-APBA)was removed from aqueous solution by a single piece of lactose-functionalized MPPM in a dynamic filtration system.The results show that the 3-APBA removal reaches an optimal efficiency(39.5%)under the alkaline condition(pH9.1),which can be improved by increasing the immobilization density of LA.Regeneration of these affinity membranes can be easily realized through acid-base washing because the complexation of boric acid and saccharide polyol is reversible.

Genetic imprinted gene PEG10 expression in deciduas of normal early pregnant women

Objective： To investigate the role and significance of paternally expressed gene 10 （PEG! 0） in deciduas of normal pregnant women. Methods： Sixty deciduas tissue from pregnant 6 to 11 weeks were divided into six groups and in each group ten samples were done every gestational week. The expression and distribution of PEG10 in deciduas were examined by RT-PCR, hybridization in situ, Western Blot and laser scanning confocal microscopy. Results： The PEG10 mRNA and protein were expressed in deciduas tissue from pregnant 6 to 11 weeks. Among them, the expression of PEG10 showed a gradual increase as the pregnancy weeks increased. In RT-PCR, the PEG10 expression was lower at pregnant 6th week （0.6743±0.114）, from pregnant 7 to 8 weeks, the expression was increased gradually （7th week 0.7349±0.0083） and reached the pinnacle at 8th week （0.7354±0.0074）. And then the pinnacle began to descend （9th week 0.6340±0.0084, 10th week 0.5901±0.0089 and llth week 0.5261±0.0112）. There was a significant difference in the expression of PEG10 from pregnant 6 to 11 weeks except 7th week and 8th week. This expression characteristic was demonstrated by hybridization in situ. The similar conclusion was proved by Western Blot and laser scanning confocal microscopy. Conclusion： Expression of PEG 10 may play an important role in early pregnancy.