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1.
Painted bilayers containing reconstituted ion channels serve as a well defined model system for electrophysiological investigations of channel structure and function. Horizontally oriented bilayers with easy solution access to both sides were obtained by painting a phospholipid:decane mixture across a cylindrical pore etched into a 200-microm thick silicon wafer. Silanization of the SiO(2) layer produced a hydrophobic surface that promoted the adhesion of the lipid mixture. Standard lithographic techniques and anisotropic deep-reactive ion etching were used to create pores with diameters from 50 to 200 microm. The cylindrical structure of the pore in the partition and the surface treatment resulted in stable bilayers. These were used to reconstitute Maxi K channels in the 100- and 200-microm diameter pores. The electrophysiological characteristics of bilayers suspended in microchips were comparable with that of other bilayer preparations. The horizontal orientation and good voltage clamping properties make the microchip bilayer method an excellent system to study the electrical properties of reconstituted membrane proteins simultaneously with optical probes.  相似文献   

2.
The objective of this study was to evaluate the bone ingrowth of a new vacuum plasma sprayed titanium surface (vps-ti) in comparison to cs-titanium implants in a g?ttinger minipig model. Fifteen g?ttinger minipigs each received the two implants, vacuum plasma sprayed titanium with a porosity of 50% and a pore size of 200 microm (vps-ti) and an implant with a similar porosity but a different pore size 500 microm (cs-ti), at the proximal femur metaphysis by press-fit technique. The pigs were euthanized at three different postsurgical periods: 4, 8 and 12 weeks. Each femur was harvested and qualitative (macroscopic and microscopic) and quantitative (histomorphometric) histological analysis was done on histological slides. The results indicated that there was a difference in bone ingrowth between the two implants, whereas the bone ingrowth of vps-ti was superior to cs-ti after 4 and 8 weeks healing time. 12 weeks post implantationem no statistiscal difference was evident. The pore size of 200 microm seemed superior to a pore size of 500 microm. Whether or not these effects lead to a better mechanical stability remains unanswered.  相似文献   

3.
Microporous polymers (with porosity up to 90%) with a well-prescribed internal microstructure were prepared in monolithic form to construct a flow-through microbioreactor in which phenol-degrading bacteria, Pseudomonas syringae, was immobilized. Initially, bacteria was forced seeded within the pores and subsequently allowed to proliferate followed by acclimatization and phenol degradation at various initial substrate concentrations and flow rates. Two types of microporous polymer were used as the monolithic support. These polymers differ with respect to their pore and interconnect sizes, macroscopic surface area for bacterial support, and phase volume. Polymer with a nominal pore size of 100 microm with phase volume of 90% (with highly open pore structure) yielded reduced bacterial proliferation, while the polymer with nominal pore size of 25 microm with phase volume of 85% (with small interconnect size and large pore area for bacterial adhesion) yielded monolayer bacterial proliferation. Bacteria within the 25 microm polymer support remained monolayered, without any apparent production of extracellular matrix during the 30-day continuous experimental period. The microbioreactor performance was characterized in terms of volumetric utilization rate and compared with the published data, including the case where the same bacteria was immobilized on the surface of microporous polymer beads and used in a packed bed during continuous degradation of phenol. It is shown that at similar initial substrate concentration, the volumetric utilization in the microreactor is at least 20-fold more efficient than the packed bed, depending on the flow rate of the substrate solution. The concentration of the bacteria within the pores of the microreactor decreases from 2.25 cells per microm2 on the top surface to about 0.4 cells per microm2 within 3 mm reactor depth. If the bacteria-depleted part of the microreactor is disregarded, the volumetric utilization increases by a factor of 30-fold compared with the packed bed. This efficiency increase is attributed to the reduction of diffusion path for the substrate and nutrients and enhanced availability of the bacteria for bioconversion in the absence of biofilm formation as well as the presence of flow over the surface of the monolayer bacteria.  相似文献   

4.
The paper presents data on the cultivation of human dermal fibroblasts and rabbit mesenchymal stromal cells on two types of porous titanium implants, i.e., those with irregular pores formed by pressed titanium particles and those with regular pores formed by the cohesion of one-size titanium particles inside the implant. The goal of this study was to determine what type of titanium implant porosity ensured its strongest interaction with cells. Cells were cultivated on implants for 7 days. During this period, they formed a confluent monolayer on the implant surface. Cells grown on titanium implants were monitored by scanning electron microscopy. Fibroblasts interaction with implants depended on the implant porosity structure. On implants with irregular pores cells were more spread. On implants with regular pores fibroblasts enveloped particles and were only occasionally bound with neighboring particles by small outgrowths. There was no tight interaction of particles inside the implant. In implants formed by pressed particles, cells grow not only on surface, but also in the depth of the implant. Thus, we suppose that a tighter interaction of cells with the titanium implant and, supposedly, tissues with the implant in the organism will take place in the variant when the implant structure is formed by pressed titanium particles, i.e., cellular interaction was observed inside the implant. In implants with irregular pores, cells grew both on the surface and in the depth. Thus, cells exhibited more adequate interactions with irregular pore titanium implants in vitro and hopefully the same interaction will be true in tissues after the implantation of the prosthesis into the organism.  相似文献   

5.
It has been reported that the activity of protein improved when it was adsorbed inside the pores of mesoporous silica (MPS). The current study investigated the activity of immobilized avidin to the biotin on MPS with various pore sizes (diameter=2.4-45.0 nm). The binding amount of immobilized avidin to biotin is 123 to 160 ng biotin/10 μg avidin on 2.7- to 5.4-nm pore MPS, but that on 12- to 45-nm pore MPS was markedly decreased (33-42 ng biotin/10 μg). Moreover, the binding amount was approximately 2- and 3-fold higher on the glycidoxypropyl (Gly)-functionalized 5.4- and 45-nm pore MPS in comparison with methyl (Me)-functionalized 5.4- and 45-nm pore MPS, respectively. Furthermore, avidin immobilized in native and Gly-grafted 45-nm pore MPS retained more than 70% and 50% binding activity to biotin, respectively, after incubating at 90°C for 3 h. In contrast, the activity was greatly reduced in the native and Gly-grafted 5.4-nm pore MPS under the same conditions (<36.9%). The immobilization also protected against effects of 0.01 M HCl and 50% MeOH; all of immobilized avidin proteins showed high activity (>50%) with biotin compared with that observed with free avidin (MeOH [<18.2%] and HCl [<32.7%]).  相似文献   

6.
Natural biodegradable polymers were processed by different techniques for the production of porous structures for tissue engineering scaffolds. Potato, corn, and sweet potato starches and chitosan, as well as blends of these, were characterized and used in the experiments. The techniques used to produce the porous structures included a novel solvent-exchange phase separation technique and the well-established thermally induced phase separation method. Characterization of the open pore structures was performed by measuring pore size distribution, density, and porosity of the samples. A wide range of pore structures ranging from 1 to 400 microm were obtained. The mechanisms of pore formation are discussed for starch and chitosan scaffolds. Pore morphology in starch scaffolds seemed to be determined by the initial freezing temperature/freezing rate, whereas in chitosan scaffolds the shape and size of pores may have been determined by the processing route used. The mechanical properties of the scaffolds were assessed by indentation tests, showing that the indentation collapse strength depends on the pore geometry and the material type. Bioactivity and degradation of the potential scaffolds were assessed by immersion in simulated body fluid.  相似文献   

7.
We report on the fabrication of an optical silicon-based label-free DNA sensor. n-Type crystalline silicon wafers have been electrochemically etched to form porous silicon layers and characterized in terms of porosity, pore distribution, surface composition and photoluminescence. Samples (0.25 cm(2)) have been cut and properly derivatized using trimethoxy-3-bromoacetamidopropylsilane in order to link single strand DNA (ss-DNA). Such a molecule is not commercially available and has been ad-hoc prepared by reacting hydrobromic acid and 3-aminopropyltrimethoxysilane in presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide as coupling agent. Trimethoxy-3-bromoacetamidopropylsilane acts as a bridge anchored to the porous silicon surface through the silane group while immobilizing ss-DNA by means of the bromoacetamido moiety. We have found that derivatized samples exhibit a photoluminescence that is stable in time and is not modified after exposure to non-complementary DNA strand. On the other hand, a sensible enhancement of the light emission has been observed when the derivatized samples react with the complementary strand, showing that the specific ss-DNA/complementary DNA (c-DNA) interaction can be optically sensed without using further labeling steps. This strongly strengthens the possible role of silicon as a material for biosensors.  相似文献   

8.
Pham QP  Sharma U  Mikos AG 《Biomacromolecules》2006,7(10):2796-2805
The physical and spatial architectural geometries of electrospun scaffolds are important to their application in tissue engineering strategies. In this work, poly(epsilon-caprolactone) microfiber scaffolds with average fiber diameters ranging from 2 to 10 microm were individually electrospun to determine the parameters required for reproducibly fabricating scaffolds. As fiber diameter increased, the average pore size of the scaffolds, as measured by mercury porosimetry, increased (values ranging from 20 to 45 microm), while a constant porosity was observed. To capitalize on both the larger pore sizes of the microfiber layers and the nanoscale dimensions of the nanofiber layers, layered scaffolds were fabricated by sequential electrospinning. These scaffolds consisted of alternating layers of poly(epsilon-caprolactone) microfibers and poly(epsilon-caprolactone) nanofibers. By electrospinning the nanofiber layers for different lengths of time, the thickness of the nanofiber layers could be modulated. Bilayered constructs consisting of microfiber scaffolds with varying thicknesses of nanofibers on top were generated and evaluated for their potential to affect rat marrow stromal cell attachment, spreading, and infiltration. Cell attachment after 24 h did not increase with increasing number of nanofibers, but the presence of nanofibers enhanced cell spreading as evidenced by stronger F-actin staining. Additionally, increasing the thickness of the nanofiber layer reduced the infiltration of cells into the scaffolds under both static and flow perfusion culture for the specific conditions tested. The scaffold design presented in this study allows for cellular infiltration into the scaffolds while at the same time providing nanofibers as a physical mimicry of extracellular matrix.  相似文献   

9.

A set of carbon monoxide (CO) gas sensors based on porous silicon (PSi)/gold nanoparticle (AuNP) hetro structures were fabricated. Different forms of PSi surface morphologies were studied as a substrate for growth of AuNPs. Simple dipping process of PSi in hydrogen tetrachloroaurate (III) solution (HAuCl4) at fixed concentrations of 10−2 M/3.5 HF was used to synthesize AuNPs. The n-type PSi was equipped through photo-electrochemical etching process at current density value of 10 mA/cm2 under illumination condition of 530-nm wavelength and laser illumination intensity of 20 to 80 mW/cm2. Three different forms of PSi morphology, meso, macro, and double layers with pore shapes and sizes, were prepared. The structural and surface morphology properties of PSi-based substrate before and after deposition of AuNPs were investigated through studying of scanning electron microscopy (SEM), photoluminescence (PL), and X-ray diffraction (XRD). The electrical property (J-V) was carried out in primary vacuum and CO at low pressure. The results show that PSi surface morphologies strongly influenced the AuNP sizes and hence the sensor performance. It was found that decrease the AuNP sizes could be occasioned in high and fast current response.

  相似文献   

10.
FORMATION AND DISTRIBUTION OF NUCLEAR PORE COMPLEXES IN INTERPHASE   总被引:4,自引:0,他引:4       下载免费PDF全文
The possibility of nuclear pore formation in the interphase nucleus was investigated in control and phytohemagglutinin (PHA) stimulated lymphocytes by the freeze-etching technique. 48 hr after the addition of PHA, the newly formed blasts which had not as yet divided had at least twice the number of pores per nucleus as controls. This clearly demonstrates that in lymphocytes nuclear pore formation can take place during interphase. It has generally been assumed that the distribution of nuclear pore complexes in somatic animal cells is random. However, we have utilized freeze etched rat kidney cells and a computer program to evaluate pore distribution. We find a minimum pore center-to-center spacing of approximately 1300 A and multiples thereof with high frequency. This is strong evidence for a nonrandom distribution of nuclear pores. The nonrandomness may be related to an underlying chromosomal organization in interphase. Using three criteria for identifying prospective pore sites (membrane specialization, nonrandomness, and alteration of heterochromatin distribution), we have found forming pores in sectioned material from cultured human melanoma cells. While nuclear pore formation may take place in conjunction with reformation of the nuclear membrane, a mechanism also exists for their formation during interphase.  相似文献   

11.
Perovskite/silicon tandem solar cells are attractive for their potential for boosting cell efficiency beyond the crystalline silicon (Si) single‐junction limit. However, the relatively large optical refractive index of Si, in comparison to that of transparent conducting oxides and perovskite absorber layers, results in significant reflection losses at the internal junction between the cells in monolithic (two‐terminal) devices. Therefore, light management is crucial to improve photocurrent absorption in the Si bottom cell. Here it is shown that the infrared reflection losses in tandem cells processed on a flat silicon substrate can be significantly reduced by using an optical interlayer consisting of nanocrystalline silicon oxide. It is demonstrated that 110 nm thick interlayers with a refractive index of 2.6 (at 800 nm) result in 1.4 mA cm?² current gain in the silicon bottom cell. Under AM1.5G irradiation, the champion 1 cm2 perovskite/silicon monolithic tandem cell exhibits a top cell + bottom cell total current density of 38.7 mA cm?2 and a certified stabilized power conversion efficiency of 25.2%.  相似文献   

12.
曹雨桐  佘冬立 《生态学杂志》2017,28(11):3684-3690
通过圆盘入渗试验,探讨不同改良剂施用量下土壤入渗特性的变化,揭示添加生物炭和聚丙烯酰胺(PAM)对海涂围垦区盐碱土水力学参数、孔隙特征及不同级别孔隙水流贡献率的影响.结果表明: 单施2%生物炭,土壤饱和导水率比对照增加46.4%;盐碱土饱和导水率随PAM施用量增加而减小.单施2%生物炭使土壤总有效孔隙度和半径>100 μm的有效孔隙度分别增加8.3%和10.2%.单施PAM时,土壤总有效孔隙度和不同半径孔隙有效孔隙度均有减小趋势,其中,PAM梯度为1‰时最明显,减幅高达88%以上.施用生物炭和PAM后,半径<100 μm的孔隙的水流贡献率呈下滑趋势,半径>500 μm的孔隙对土壤水流运动起主导作用.  相似文献   

13.
The templating approach is a powerful method for preparing porous electrodes with interconnected well‐controlled pore sizes and morphologies. The optimization of the pore architecture design facilitates electrolyte penetration and provides a rapid diffusion path for lithium ions, which becomes even more crucial for thick porous electrodes. Here, NaCl microsize particles are used as a templating agent for the fabrication of 1 mm thick porous LiFePO4 and Li4Ti5O12 composite electrodes using spark plasma sintering technique. These sintered binder‐free electrodes are self‐supported and present a large porosity (40%) with relatively uniform pores. The electrochemical performances of half and full batteries reveal a remarkable specific areal capacity (20 mA h cm?2), which is 4 times higher than those of 100 µm thick electrodes present in conventional tape‐casted Li–ion batteries (5 mA h cm?2). The 3D morphological study is carried out using full field transmission X‐ray microscopy in microcomputed tomography mode to obtain tortuosity values and pore size distributions leading to a strong correlation with their electrochemical properties. These results also demonstrate that the coupling between the salt templating method and the spark plasma sintering technique turns out to be a promising way to fabricate thick electrodes with high energy density.  相似文献   

14.
This experimental study on laser-textured implants aimed to evaluate periimplant bone elasticity and ultimate stress of the bone-implant interface in a rabbit femur model. After randomization, two cylindrical Ti6Al4V samples (3.5 mm wide, 5.5 mm long) were transcortically implanted in each femur of 15 female New Zealand White Rabbits. Polished implants had been laser-textured with 100, 200, and 300 microm diameter pores, and another corundum blasted implant was additionally textured with 200 microm pores. Twelve weeks into the experiment, a modified push-out test was performed. The median shear modulus indicating the elasticity of the periimplant bone was 41.12 MPa for the proximal implant location and 25.38 MPa for the distal, without evidence for significant differences between implant types. Taking into account the median ultimate shear stress for 200 microm implants with and without corundum blasting, no significant difference could be demonstrated. However, for blasted 200 microm implants a statistically significant (p<0.025) relative gain in ultimate shear stress of 41% and 17% was proven in comparison with 100 and 300 microm implants, respectively. Non-blasted 200 microm implants reached 48% relative gain in respect of 100 microm samples.  相似文献   

15.
Mycobacteria protect themselves with an outer lipid bilayer, which is the thickest biological membrane hitherto known and has an exceptionally low permeability rendering mycobacteria intrinsically resistant against many antibiotics. Pore proteins mediate the diffusion of hydrophilic nutrients across this membrane. Electron microscopy revealed that the outer membrane of Mycobacterium smegmatis contained about 1000 protein pores per microm(2), which are about 50-fold fewer pores per microm(2) than in Gram-negative bacteria. The projection structure of the major porin MspA of M. smegmatis was determined at 17 A resolution. MspA forms a cone-like tetrameric complex of 10 nm in length with a single central pore. Thus, MspA is drastically different from the trimeric porins of Gram-negative bacteria and represents a new class of channel proteins. The formation of MspA micelles indicated that the ends of MspA have different hydrophobicities. Oriented insertion of MspA into membranes was demonstrated in lipid bilayer experiments, which revealed a strongly asymmetrical voltage gating of MspA channels at -30 mV. The length of MspA is sufficient to span the outer membrane and contributes in combination with the tapering end of the pore and the low number of pores to the low permeability of the cell wall of M. smegmatis for hydrophilic compounds.  相似文献   

16.
Zhu X  Cui W  Li X  Jin Y 《Biomacromolecules》2008,9(7):1795-1801
Diffusional limitations of mass transport have adverse effects on engineering tissues that normally have high vascularity and cellularity. The current electrospinning method is not always successful to create micropores to encourage cell infiltration within the scaffold, especially when relatively large-sized pores are required. In this study, a slow rotating frame cylinder was developed as the collector to extend the pore size and increase the porosity of electrospun fibrous scaffolds. Fibrous mats with porosity as high as 92.4% and average pore size of 132.7 microm were obtained. Human dermal fibroblasts (HDFs) were seeded onto these mats, which were fixed on a cell-culture ring to prevent the shrinkage and contraction during the incubation. The viability test indicated that significantly more HDFs were generated on highly porous fibrous mats. Toluidine blue staining showed that the highly porous scaffold provided mechanical support for cells to maintain uniform distribution. The cross-section observations indicated that cells migrated and infiltrated more than 100 microm inside highly porous fibrous mats after 5 d incubation. The immunohistochemistry analysis demonstrated that cells began secreting collagen, which is the main constituent of extracellular matrix. It is supposed that highly porous electrospun fibrous scaffolds could be constructed by this elaboration and may be used for skin tissue engineering.  相似文献   

17.
The tensile strength and stiffness of load-bearing soft tissues are dominated by their collagen fiber orientation. While microgrooved substrates have demonstrated a capacity to orient cells and collagen in monolayer tissue culture, tissue engineering (TE) scaffolds are structurally distinct in that they consist of a three-dimensional (3-D) open pore network. It is thus unclear how the geometry of these open pores might influence cell and collagen orientation. In the current study we developed an in vitro model system for quantifying the capacity of large scale ( approximately 200 microm), geometrically well-defined open pores to guide cell and collagen orientation in engineered tissues. Non-degradable scaffolds exhibiting a grid of 200 microm wide rectangular pores (1:1, 2:1, 5:1, and 10:1 aspect ratios) were fabricated from a transparent epoxy resin via high-resolution stereolithography. The scaffolds (n=6 per aspect ratio) were surface modified to support cell adhesion by covalently grafting GRGDS peptides, sterilized, and seeded with neonatal rat skin fibroblasts. Following 4 weeks of static incubation, the resultant collagen orientation was assessed quantitatively by small angle light scattering (SALS), and cell orientation was evaluated by laser confocal and scanning electron microscopy. Cells adhered to the struts of the pores and proceeded to span the pores in a generally circumferential pattern. While the cell and collagen orientations within 1:1 aspect ratio pores were effectively random, higher aspect ratio rectangular pores exhibited a significant capacity to guide global cell and collagen orientation. Preferential alignment parallel to the long strut axis and decreased spatial variability were observed to occur with increasing pore aspect ratio. Intra-pore variability depended in part on the spatial uniformity of cell attachment around the perimeter of each pore achieved during seeding. Evaluation of diamond-shaped pores [Sacks, M.S. et al., 1997. J. Biomech. Eng. 119(1), 124-127] suggests that they are less sensitive to initial conditions of cell attachment than rectangular pores, and thus more effective in guiding engineered tissue cell and collagen orientation.  相似文献   

18.
A label-free immunosensor for the detection of α-fetoprotein (AFP) is proposed based on controlled fabrication of monoclonal antibodies of AFP (anti-AFP) and gold nanoparticles (GNPs) inside the pores of mesoporous silica (MPS). The silanol groups on the internal pore walls were grafted by aminopropyltriethoxyl silane, whereas the silanol groups on the external surface of MPS were blocked by trimethylchlorosilane (TMCS). Thus, anti-AFP and GNPs could be confined inside the mesopores of TMCS-MPS by the covalent linking with the amino groups. The prepared anti-AFP/GNPs/TMCS-MPS particles were used to modify glassy carbon electrode (GCE) to construct a label-free immunosensor. After incubating the sample AFP with the anti-AFP/GNPs/TMCS-MPS/GCE, the immunoconjugates were formed on the surface of GCE and the spatial block increased. Thus, the peak current decreased with increasing concentrations of AFP. GNPs inside the mesopores could promote the electron transportation through the pore channel. Under the optimal experimental conditions, the fabricated immunosensor could detect AFP in a linear range from 1.0 to 90 ng ml(-1) with a detection limit of 0.2 ng ml(-1) (3σ). It provided a novel alternative method for the label-free determination of other antigens.  相似文献   

19.
A numerical model of a human body with an intramedullary nail in the femur was built to evaluate the effects of the implant on the current density distribution in extremely low frequency electric and magnetic fields. The intramedullary nail was chosen because it is one of the longest high conductive implants used in the human body. As such it is expected to alter the electric and magnetic fields significantly. The exposure was a simultaneous combination of inferior to superior electric field and posterior to anterior magnetic field both alternating at 50 Hz with the values corresponding to the ICNIRP reference levels: 5000 V m?1 for electric field and 100 µT for magnetic flux density. The calculated current density distribution inside the model was compared to the ICNIRP basic restrictions for general public (2 mA m?2). The results show that the implant significantly increases the current density up to 9.5 mA m?2 in the region where it is in contact with soft tissue in the model with the implant in comparison to 0.9 mA m?2 in the model without the implant. As demonstrated the ICNIRP basic restrictions are exceeded in a limited volume of the tissue in spite of the compliance with the ICNIRP reference levels for general public, meaning that the existing safety limits do not necessarily protect implanted persons to the same extent as they protect people without implants. Bioelectromagnetics 30:591–599, 2009. © 2009 Wiley‐Liss, Inc.  相似文献   

20.
冻融交替后不同尺度黑土结构变化特征   总被引:10,自引:0,他引:10  
王恩姮  赵雨森  夏祥友  陈祥伟 《生态学报》2014,34(21):6287-6296
冻融交替是改变黑土结构、加剧土壤侵蚀的重要因子。以典型黑土区耕作土壤为研究对象,采用野外季节性冻融循环与室内模拟冻融循环相结合、X射线计算机断层摄影(CT)与扫描电子显微镜(SEM)相结合的方法,通过水分物理性质、团聚体破坏率、孔隙数目、孔隙面积、孔隙成圆率、孔隙Feret直径的测定与分析,研究了冻融交替后0—40 cm、40—80 cm和120—160 cm3个土层以及田间季节性冻融环刀、室内模拟冻融CT扫描和室内模拟冻融SEM3种方式下黑土结构特征的变化规律。结果表明:冻融交替能够对不同土层和不同尺度的耕地黑土结构产生不同程度的影响。季节性冻融后,表层土壤容重升高,非毛管孔隙度和持水能力显著降低(P0.05),40—80 cm土层团聚体破坏率增加40.97%(P0.05),土壤抗蚀性有所削弱,120—160 cm土壤没有受到季节性冻融的显著影响。CT扫描尺度上,3个土层均以1—2 mm径级的孔隙数目为最多,形状也相对规则、接近圆形;冻融循环没有对表层土壤大孔隙结构产生影响,却能够显著降低40—80 cm土层范围内大孔隙面积以及Feret直径(P0.05)。SEM扫描显示冻融后土壤表面粗糙度增加,颗粒松散、脱离,孔壁断裂,证明了冻融交替对土壤微结构的破坏作用;同时结合电子能谱的元素分析可知冻融交替能够改变土壤颗粒表面化学特征。  相似文献   

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