Ratio Of Staining Area Research Articles

Study on Plant-blanket to reduce heavy metal migration caused by precipitation and to improve the soil environment of pyritic tailings

The increasing demand for mineral resources due to industrial development has led to significant tailings pollution during the mineral extraction process. In the southwestern region of China, a large amount of pyritic tailings containing pyrite cinder easily leaches heavy metals and other pollutants when exposed to precipitation, resulting in widespread soil contamination. Effective remediation methods are urgently needed to address this issue. This study utilized naturally occurring Plant-blanket formed by the symbiosis of moss and herbaceous plants on pyritic tailings as restoration material. Through leaching experiments and staining tracer techniques, the study investigated the ability of Plant-blanket to reduce the migration of heavy metals from pyrite cinder to soil under the influence of precipitation and its role in improving the soil environment. The results showed that within 12 h, the Plant-blanket could absorb water equivalent to 206.9 % of its own weight and had good water retention ability. It reduced the stained area ratio of soil horizontal and vertical profiles after precipitation leaching by a maximum of 76.08 % and 46.41 %, respectively, and improved the pH, cation exchange capacity (CEC), bulk density, and water content of soil at different depths. In addition, after being covered by Plant-blanket, the migration of Cd and Cu was reduced by a maximum of 44.35 % and 55.77 % respectively, and it increased the diversity and abundance of bacterial communities, promoting the recovery of soil microbial ecological functions. These findings indicate that Plant-blanket can regulate water and improve soil environment, and has certain control ability on the migration of Cd and Cu produced by pyritic tailings. Meanwhile, Plant-blanket plays an important role in improving the soil environment in mining areas and promoting ecosystem restoration, providing valuable reference for further exploration of ecological restoration of tailings.

Impacts of moss-dominated biocrusts on rainwater infiltration, vertical water flow, and surface soil evaporation in drylands

Globally widespread biocrusts form a “living skin” on most dryland soils, helping govern fundamental ecosystem functions, particularly hydrological processes. However, the underlying mechanisms of biocrust effects on soil water infiltration and evaporation are still controversial. Biocrusts possibly regulate infiltration and evaporation due to their unique physicochemical properties and influence on surface soil moisture (θ), and these assumptions were verified in this study conducted on the Loess Plateau of China. The differences in vertical soil moisture distribution and flow patterns of infiltration water between moss-dominated biocrusts and bare soil were investigated through dye tracing experiments. The evaporation characteristics of biocrusts and bare soil were also obtained by weighing micro-lysimeters in both simulated and in-situ experiments. Results showed that biocrusts retarded deep soil water infiltration below ∼ 15 cm and significantly (F = 48.15, P < 0.001) decreased the stained area ratio (percentage of stained area to total area at the same soil depth) by 16% in contrast to bare soil. Because the infiltrated water was mostly retained in shallow soil (0–15 cm) in biocrust-covered soil, their ratios of low interaction flow (stained path width < 20 mm) and high interaction flow (20 mm < stained path width < 200 mm) were significantly higher (F ≥ 7.82, P ≤ 0.007) than those of bare soil. After the dye tracing experiments, the biocrusts modestly increased θ by 17% throughout the soil profile (0–50 cm), but they remarkably increased θ by 33%−78% in the top 20 cm of soil. Additionally, the biocrusts increased soil daily evaporation rate (E) by 27%−39% (with only the exception of the beginning) and the cumulative evaporation amount by 14% in contrast to bare soil over an 8-day field measurement. More importantly, the biocrust-affected soil properties, such as the contents of fine particles and organic matter, as well as saturated water content and field capacity, were closely correlated with E (|ρ| ≥ 0.52, P ≤ 0.08). Consequently, it was concluded that biocrusts restrict the depth of rainwater infiltration and increase surface θ, which may greatly enhance surface soil evaporation in the Chinese Loess Plateau.

Water seepage and retention in purple soil bunds on sloping farmland in the Three Gorges Reservoir area, China

AbstractSoil bunds are a common land reconstruction practice in the Three Gorges Reservoir area of China, and play a key role in runoff regulation and soil conservation on cultivated hillslopes. This study investigated water movement pathways and retaining effects of soil bunds using water transport trajectory observations inferred from field dye tracing experiments. It was found that water infiltration in the bund was characterised by spatial heterogeneity in three dimensions. The stained area ratio (SAR) of the bund was much lower than that of the adjacent field at all depths. Dye patterns resembled inclined strips in the bund, and lateral seepage occurred mainly on the top of hard pan. SAR, stained path width (SPW) &lt;20 mm, and stained path number (SPN) all decreased with increasing horizontal distance. The SPW &lt;20 mm proportion and SAR both decreased according to a logarithmic function, while the changing trend of SPN followed a first‐order exponential decay. Three pathways of water movement in the bund could be distinguished, namely, vertical infiltration, lateral seepage and flow along the bund–bedrock interface. The primary water seepage channel in the bund was likely to be the bund–bedrock interface (accounting for around 80% of the total flow). These results provide preliminary realisation of water movement mechanisms in the purple soil bunds on sloping farmland and inform management options for bund stability maintenance.

Effects of straw returning to movement characteristics of soil preferential flow in sugarcane fields

The measures of returning straw to the field can change soil environment in the field and affect crop growth. In order to explore the impacts of different straw returning measures on the preferential flow in the field, we investigated the movement characteristics of preferential flow in sugarcane fields under different straw returning measures [straw mulching (CM), straw burning mulching (BM), and non-straw mulching (CK)] in Guangxi by combining image analysis techniques with ecological landscape pattern analysis methods. The results showed that under the same external water supply environment, soil staining morphology of non-straw mulching sugarcane fields differentiated into finger-like and clump-like. Both straw mulching and straw burning mulching were mainly differentiated into clump-like. The mean total staining area ratio was significantly higher than that of non-straw mulching (26.0%). The ratio of surface staining area decreased rapidly under non-straw mulching. The water movement range decreased rapidly under straw mulching and straw burning mulching in the depth of 15-30 cm. The change rate of straw mulching staining area ratio was the lowest in the deep soil. The shape index of non-straw mulching was 15.54. The water flow movement was more concentrated. The average sub-circular index of straw mulching and straw burning mulching was 0.67 in the sugarcane field, with a higher degree of preferential flow bending, an average proximity index of 0.87, and better water flow connectivity. The average substrate flow depth (3.52 cm) and edge density index (11.51) were the smallest under non-straw mulching, but the priority flow ratio was the largest (73.2%), and the spatial development degree of preferential flow was the highest. The straw returning measures reduced the preferential flow in the sugarcane fields, with positive effects on water conservation.

A Study of Plaque Adhesion during Implant Treatment of Missing Molars

Purpose: In implant treatment, the abutments and superstructures form a transmucosal area. In addition to providing appropriate cleaning instructions to patients, its material properties must be carefully considered. It is not clear how much plaque adhesion occurs at each site during treatment process, and the effect of materials on plaque adhesion. Methods: In this study, 33 patients, 13 maxillary cases and 20 mandibular cases, who had implants placed in the missing molars were evaluated for the plaque adhesion of provisional restorations made of acrylic resin and monolithic zirconia final restoration after a period of more than 4 weeks. In addition, oral hygiene instructions were thoroughly given for 12 cases at the time of staining after the provisional restorations, and re-evaluated in 4 weeks. Results: About 20% of the buccal-lingual area of the prosthetic device fabricated with acrylic resin showed plaque staining, whereas about 40% of the area of the mesial and distal. On the other hand, for the zirconia final restorations, the percentage of staining was about 5% for buccal and lingual surface and 10% for mesial and distal surface. The stained area ratio of the provisional restoration correlated with the zirconia superstructure area ratio. Even with careful oral hygiene, it became clear that the amount of plaque adhesion due to the difference in materials could not be overcome. Conclusion: The zirconia superstructure is extremely good from the viewpoint of plaque adhesion, but cases with a large stained area at the time of provisional restoration should be carefully followed up.

Determining the influencing factors of preferential flow in ground fissures for coal mine dump eco-engineering.

Ground fissures (GF), appearing in front of dumps, are one of the most obvious and harmful geological hazards in coal mining areas. Studying preferential flow and its influencing factors in the ground fissures of dumps may provide basic scientific support for understanding the rapid movement of water and vegetation restoration and reconstruction in mining areas. Based on field surveys of ground fissures, three typical ground fissures were selected in the studied dump. The morphological characteristics of preferential flow for ground fissures were determined through field dye tracing, laboratory experiments, and image processing technology. The results indicated that the lengths of the three ground fissures ranged from 104.84 cm to 120.83 cm, and the widths ranged from 2.86 cm to 9.85 cm. All of the ground fissure area densities were less than 10%, and the proportion of ground fissure surface area was small in the dump. The maximum fissure depth was 47 cm, and the minimum was 16 cm. The ground fissure widths ranged from 0 cm to 14.98 cm, and the fissure width and fissure width-to-depth ratios decreased with increasing soil depth. The stained area was greater than 90% in the 0–5 cm soil layers of the three fissures, and water movement was dominated by matrix flow. The stained width decreased from 90 cm to 20 cm with increasing soil depth. The preferential flow was mainly concentrated on both sides of the fissure, which was distributed as a “T” shape. The preferential flow stained area ratios were 27.23%, 31.97%, and 30.73%, respectively, and these values decreased with increasing soil depth. The maximum stained depths of the preferential flow among the three fissures were different, and the maximum stained depth of GF II was significantly larger than that of GF I and GF III (P < 0.05). The stained path numbers of the three fissures ranged from 0 to 49. With increasing soil depth, the stained path number first increased and then decreased. The stained path widths of the three fissures ranged from 0 cm to 90 cm. With the increase in soil depth, the stained path width decreased. The stained area ratio was significantly positively correlated with ground fissure width, the ground fissure width-to-depth ratio, soil saturated hydraulic conductivity, soil organic matter, and sand content and was significantly negatively correlated with soil water content and clay content. The stained path number was significantly positively correlated with ground fissure width, the ground fissure width-to-depth ratio, soil saturated hydraulic conductivity and soil organic matter. The stained path width was significantly positively correlated with the ground fissure width-to-depth ratio, soil saturated hydraulic conductivity, soil organic matter and sand content and was significantly negatively correlated with clay content. Plant roots could significantly increase the stained area ratio, stained path number, and width and promote the formation and development of preferential flow.

Apoptosis in the liver of male &lt;em&gt;db/db&lt;/em&gt; mice during the development of obesity and type 2 diabetes

Obesity and diabetes mellitus are known to lead to the development of metabolic syndrome and non-alcoholic fatty liver disease (NAFLD). The mechanisms of programmed cell death are actively involved in maintaining cellular homeostasis along development of NAFLD. Proteins of the BCL-2 family are key regulators of physiological and pathological apoptosis. Homozygous males of BKS.Cg-Dock7mLeprdb/+/+/J mice (db/db mice) are characterized by progressive obesity and the development of type 2 diabetes mellitus (DM2) with severe hyperglycemia at 4–8 weeks and organ lesions at 8–10 weeks of age. The aim of this research was to study the expression of molecular cell regulators of apoptosis in liver cells of db/db mice males at different stages of obesity and diabetes development (at the age of 10 and 18 weeks). Immunohistochemical analysis (using the indirect avidin-biotin peroxidase method) and morphometric evaluation of the expression of the antiapoptotic protein Bcl-2 and the proapoptotic protein Bad in liver cells of studied animals at different stages of obesity and DM2 were carried out. An excess of the value of the Bcl-2 protein staining area over the Bad protein staining area was revealed in the liver of 10-week-old animals. The Bcl-2/Bad expression area ratio in 10-week-old animals was twice as high as in 18-week-old animals, which indicates the presence of conditions for blocking apoptosis in the liver of younger db/ db mice. At the 18th week of life, db/db mice displayed an almost threefold increase in the expression area of the Bad protein against the background of an unchanged expression of the Bcl-2 protein. The decrease in the Bcl-2/Bad staining area ratio in 18-week-old animals was due to the increase in the Bad expression area, which indicates the absence of antiapoptotic cell protection and creates conditions for activation of the mitochondrial pathway of apoptosis in the liver of male db/db mice with pronounced signs of obesity and DM2.

Effects of cultivation history in paddy rice on vertical water flows and related soil properties

Soil properties, especially physical properties and macropore characteristics, are controlling factors for vertical water flow in paddy fields. However, the effects of paddy rice cultivation history on vertical water flows and related soil properties have been rarely studied. The objectives were to identify the differences in soil physical properties and macropore parameters and to reveal the effects of these soil properties on vertical water flow patterns in paddy fields under different cultivation times. In this study, soil physical properties in a very young paddy field (2 y, VYPF), a young paddy field (18 y, YPF), and an old paddy field (>100 y, OPF) were analyzed. Meanwhile, macropore parameters (resolution of 0.6 mm) and vertical water flow patterns in these fields were identified by computed tomography and dye tracing experiment, respectively. As cultivation history lengthened in paddy rice, the average content of soil clay and organic carbon in the 80 cm depth soil profile increased, as did the thickness and bulk density of the plow pan. Also, macroporosity, macropore area density, macropore number density, node density, macropore angle, and hydraulic radius significantly decreased as length of cultivation age increased. As a result, differences in these soil propeties affected vertical water flow in three studied paddy fields. Steady-state infiltration rates decreased significantly in accordance with the reduced saturated hydraulic conductivity of soil cores from the plow pan; median values were 9.76 cm d−1, 2.93 cm d−1, and 0.13 cm d−1 for VYPF, YPF, and OPF, respectively. A greater abundance and distribution of a dye tracer and a higher stained area ratio under the plow pan in younger paddy fields indicated that more obvious preferential water flows occurred in VYPF and YPF than in OPF. This implied that the conversion of farmland from upland field to paddy field resulted in greater use of irrigation water and also posed a risk of groundwater pollution through increased percolation. Therefore, a longer puddling time before planting paddy rice should be conducted in paddy fields (i.e. YPF and VYPF) with a short cultivation history, which can reduce water percolation by enhancing bulk density, reducing macropores, and lowering the saturated water conductivity of the plow pan.

Renal elasticity and perfusion changes associated with fibrosis on ultrasonography in a rabbit model of obstructive uropathy.

To evaluate elasticity and perfusion change associated with fibrosis in a rabbit model of unilateral ureter obstruction using shear wave elastography (SWE) and contrast-enhanced ultrasonography (CEUS). Complete unilateral ureter obstruction by ligation was performed in the left kidney of 15 rabbits. Renal elasticity on SWE and perfusion change on CEUS at the renal cortex were measured before and after the operation. Histopathological renal fibrosis was quantified by the stained area ratio with Masson trichrome and Picrosirius red using ImageJ analysis. Renal elasticity and perfusion values were compared by the Mann-Whitney U test and Proc Mixed as a function of time. Spearman's correlation was used to analyze differences between imaging values and fibrosis. The duration of imaging follow-up was up to 49 days, with interval imaging performed 1-3 times. Renal elasticity values were higher in obstructed kidneys compared to contralateral kidneys (31.0 kPa vs 16.4 kPa, p < 0.001) and increased according to postoperative time (0.46 kPa/day). With respect to renal fibrosis, SWE values were positively correlated with Masson trichrome (ρ = 0.651, p < 0.001) and Picrosirius red (ρ = 0.514, p = 0.007). Among CEUS parameters, mean transit time was negatively correlated with renal fibrosis by Masson trichrome (ρ = - 0.639, p = 0.001) and Picrosirius red (ρ = - 0.625, p = 0.001). Rise time and time to peak were positively correlated with renal fibrosis. Obstructive uropathy resulted in changes to both renal elasticity and perfusion. Renal fibrosis was moderately associated with increased renal cortical stiffness and both delayed and decreased cortical perfusion. • Obstructive uropathy causes changes in elasticity and perfusion in a rabbit model. • Renal fibrosis from obstructive uropathy increases renal cortical stiffness, and both delay and decrease cortical perfusion.

HISTOLOGICAL OF CHANGES IN THE AMNIOTIC MEMBRANE AND PLACENTAL VILLOUS BASAL LAMINA IN COMPLICATED PREGNANCIES

Aim: This study aimed to histological changes compare placental villous basal lamina and amniotic membrane changes in complicated pregnancy. Methods : Studies were performed on the human placentas, delivered between 24-39 weeks of gestation. Patients were separated equally into 4 groups ( Control, preeclampsia (PE), gestastional diabetes (GD), and HELLP syndrome groups ) Placental tissue samples were dissected and fixed in 10% neutral formalin buffer. Routine paraffin tissue protocol was followed. Some of sections were stained with Periodic Acid Schiff. Remaining sections were stained with integrin alpha-6 antibody.To define expression percentage, mean of the staining area/total staining area ratio were calculated. The statistical significance of the expression percentages was compared by One Way ANOVA and Tukey tests with SPSS Statistics V24 software. Results: In PAS-stained preeclamptic, HELLP and gestational diabetes groups placental villous basal lamina and vasculo-syncytial membranes were thicker than the control group. A significant difference was observed in all 3 groups compared to the control group the placental villous basal lamina thickness of the HELLP group was found to be significantly different from all three groups. In chorionic villi of HELLP group, dense integrin expression was found in placental villous basal lamina similar to that in GD and preeclampsia groups. The HELLP group was significantly different from all groups. Conclusion: In preeclampsia, gestational diabetes and HELLP placentas, placental viioz basal lamina and amniotic membrane significantly thickened and structural changes were observed.

Changes of preferential flow path on different altitudinal zones in the Three Gorges Reservoir Area, China

Liu, M., Du, W. and Zhang, H. 2014. Changes of preferential flow path on different altitudinal zones in the Three Gorges Reservoir Area, China. Can. J. Soil Sci. 94: 177–188. Preferential flow in soil macropores plays an important role in runoff control and soil and water conservation. The aim of this study was to investigate the distribution of preferential flow paths in the soil profile of various altitudinal belts, analyze its variation among different soil horizons, and define the cause of soil macropores. A dye tracer method combined with photographic analysis was conducted for four hillslope sites in the Three Gorges Reservoir Area of China (TGRA). The results show that stained area proportion, as well as its vertical distribution in soil sections, presented varied patterns due to changes of forest vegetation and soil type with altitude. Stained area ratio of soil profiles increased, while stained depth decreased with increasing altitude. For soil sections in the subalpine belt, mid-mountain belt, and low-mountain belt of TGRA, stained area ratios were 62, 42, and 45%, and stained depths were 52.4, 56.4, and 69.5 cm, respectively. For brown earth covered with subalpine temperate deciduous broad-leaved forest, stained area ratios were the largest, but dyed patches were concentrated in the humus horizon. For yellow earth covered with low-mountain warm coniferous forest, stained depth reached 69.5 cm, and stained patches existed in the total soil profile. Compared with forest soil, stained depth and stained area ratio of abandoned farmland in low-mountain belt were lower, and the depth of dye infiltration was even shallower.

Changes of preferential flow path on different altitudinal zones in the Three Gorges Reservoir Area, China

Liu, M., Du, W. and Zhang, H. 2014. Changes of preferential flow path on different altitudinal zones in the Three Gorges Reservoir Area, China. Can. J. Soil Sci. 94: 177-188. Preferential flow in soil macropores plays an important role in runoff control and soil and water conservation. The aim of this study was to investigate the distribution of preferential flow paths in the soil profile of various altitudinal belts, analyze its variation among different soil horizons, and define the cause of soil macropores. A dye tracer method combined with photographic analysis was conducted for four hillslope sites in the Three Gorges Reservoir Area of China (TGRA). The results show that stained area proportion, as well as its vertical distribution in soil sections, presented varied patterns due to changes of forest vegetation and soil type with altitude. Stained area ratio of soil profiles increased, while stained depth decreased with increasing altitude. For soil sections in the subalpine belt, mid-mountain belt, a...

The correlation between extracellular resistance by electrical biopsy and the ratio of optical low staining area in irradiated intestinal tissues of rats

BackgroundElectrical biopsy illustrates a tissue’s electrical properties by electrical impedance spectroscopy. However, electrical biopsy parameters are different from conventional morphological-based examinations. The correlation between electrical biopsy and the morphological observation has not been checked. Considering the tissue responses to injury, extracellular resistance should be most sensitive with the accumulation of fluid in tissue, and it is expected to increase the ratio of optical low staining area on histological images. In this study, we calculated the ratio of optical low staining area of sampled histological images and compared with the results of electrical biopsy to verify the hypothesis of that the extracellular resistance of electrical biopsy most highly correlates with the ratio of optical low staining area on histological images.MethodsThe irradiated intestinal tissues of rats after different latent period were used for study. The sampled tissues were measured by electrical impedance spectroscopy for electrical biopsy and the microscopic images were acquired. The sampled histological images were transformed into the Hue-Saturation-Density (HSD) colour model to decouple the stain density. The ratio of optical low staining area on histological images was computed to quantify the morphological changes. The results were related to the parameters from electrical biopsy according to three element circuit model by Spearman’s rank correlation test.ResultsThe ratio of optical low staining area varied as well as the tissue’s electrical parameters. The extracellular resistance (Re) and intracellular resistance (Ri) by electrical biopsy tended to increase with the ratio of low staining area decreasing. The membrane capacitance (Cm) by electrical biopsy tended to increase with the ratio of optical low staining area increasing. The extracellular resistance (Re) of electrical biopsy was the parameter most highly correlated with the ratio of optical low staining area with a correlation coefficient of −0.757 (p < 0.001).ConclusionsThe results of this report confirm the hypothesis and support the idea that electrical biopsy results reflect the changes in tissues seen in conventional histological findings in a sense of conventional histological knowledge, and this approach may have a great potential for augmenting the pathological diagnosis of tissues.

Effects of selective phosphodiesterase 4 inhibitor on expression of aquaporin 5 in airway mucus hypersecretion model of rats

To explore the mechanism of selective phosphodiesterase 4 (PDE4) inhibitor and observe its effects on the in vitro expression of aquaporin 5 (AQP5) in a rat model of airway mucus hypersecretion. Forty healthy male rats were randomized into 4 groups (n = 10 each). The normal control and drug control groups underwent normal saline aerosol inhalation or YM976 lavage respectively for 12 days while the model and treatment groups acrolein atomization inhalation or acrolein atomization inhalation plus YM976 lavage respectively for 12 days. The middle lobe of right lung were fixed, embedded and stained by hematoxylin and eosin to observe the histological changes of small airway. The mucin secretion by goblet cells in lung tissue was tested by alcian blue staining. And the effects of selective PDE4 inhibitor on the expression of AQP5 were detected by immunohistochemical stain. Reverse transcription-polymerase chain reaction and Western blot were used to determine the levels of AQP5 mRNA and protein. In the treatment group, mucus gland hyperplasia and airway mucus hypersecretion and positive staining area ratio of alcian blue staining decreased significantly compared with the model group (10.23% ± 0.94% vs 14.74% ± 1.06%, P < 0.05). And the expression of AQP5 mRNA (1.26 ± 0.19) and protein (0.56 ± 0.13) also declined significantly versus the model group ((0.96 ± 0.17), (0.43 ± 0.15), P < 0.05). Selective PDE4 inhibitors alleviate the pathological damage of lung tissue and enhance the expression of AQP-5 in airway mucus hypersecretion.

Repair of ulcer with rhEGF sustained-release microspheres in diabetic rats

Objective To prepare recombinant human epidermal growth factor (rhEGF) sustained-release microspheres and evaluate their morphology, rhEGF releasing activities and cell proliferation activity in vitro and compare difference of rhEGF sustained-release microspheres and rhEGF in facilitaring ulcer healing in diabetic rats. Methods (1) rhEGF sustained-release microspheres were prepared by the modified double emulsion method. Morphology of the microspheres was detected by transmission electron microscope and size distribution measured by laser granularity meter/Zeta electric potential meter. ELISA assays were applied to determine rhEGF releasing. (2)Proliferation of mouse fibroblasts was analyzed by MTr method. (3) Diabetic rat models were prepared and divided into four groups, ie, rhEGF sustained-release mierospheres group (Group A), rhEGF stock solution group (Group B), blank sustainedrelease mierospheres group (Group C) and PBS meustruum control group (Group D), which were given drug once a day. The wound healing rate was calculated by taking photographs at days 3,7,14 and 21. Skin specimens from the wound edge were harvested partially for observation of hydroxyproline (HYP) contents. Immunohistochemistry was employed to detect integrin 131 and keratin-19 and measure their positive staining area ratio. Results (1) The particle diameter of rhEGF sustained-release microspheres was 193.5 nm, with relative uniform particle diameter distribution. There showed no conglutination among rhEGF susrained-release microspheres, with good dispersibility. Releasing drug lasted for 24 hours and accorded with Higuchi release kinetic model. (2) Different concentrations of rhEGF sustained-release microspheres could promote the proliferation of mouse fibroblast, especially the concentration of 10 μg/L (P <0.05, compared with the control). (3) From the 7th day after treatment, Group A had the fastest wound healing rate, with statistical difference compared with other three groups (P < 0.05). Group A had higher HYP contents and positive area ratio of integrin β1 and keratin-19 than Group B. Conclusions rhEGF sustained-release microspheres prepared by the modified double emulsion method have uniform particle size and can last release for 24 hours. Compared with rhEGF stock solution, rhEGF sustained-release microspheres have faster and better ulcer healing and higher healing quality in diabetic rats. Key words: Diabetes mellieus; experimental; Skin ulcer; Nanostructures; Epidermal growth factor; Hydroxyproline; Antigen; CD29; Keratin-19

Difference between Deep Seawater and Surface Seawater in the Preventive Effect of Atherosclerosis

Using surface and deep seawater collected in the sea area of Muroto Cape (Kochi, Japan), desalinated drinking samples of about 1200 hardness were prepared and examined for the effects on the prevention of atherosclerosis in dietary induced hyperlipidemia rabbits. The plasma LDL cholesterol level was lower in the deep seawater group than in the surface seawater group. GPx activity was significantly higher in the deep seawater group than in the control group, while there was no difference between the surface seawater and control groups. The level of LPO was also significantly lower in the deep seawater group than in the control group. The Sudan IV lipid stained area ratio on the inner surface of the aorta was significantly lower in the deep seawater groups than in the control group, while there was no difference between the surface seawater and control groups. The oil red O stained cross section of the aorta in the control and surface seawater administration group foam cells had accumulated to form thick layers, while in the deep seawater administration group, the degree of their accumulation was very low. These results suggested that the deep seawater was useful for the prevention of hyperlipidemia and arteriosclerosis compared to the surface seawater, and it was found that reduction of the LDL cholesterol level and enhancement of GPx activity were involved in its effects.

Enhanced expression of fibroblast growth factors and receptor FGFR-1 during vascular remodeling in chronic obstructive pulmonary disease.

Important characteristics of chronic obstructive pulmonary disease (COPD) include airway and vascular remodeling, the molecular mechanisms of which are poorly understood. We assessed the role of fibroblast growth factors (FGF) in pulmonary vascular remodeling by examining the expression pattern of FGF-1, FGF-2, and the FGF receptor (FGFR-1) in peripheral area of lung tissues from patients with COPD (FEV(1) < or = 75%; n = 15) and without COPD (FEV(1) > or = 85%; n = 13). Immunohistochemical staining results were evaluated by digital video image analysis as well as by manual scoring. FGF-1 and FGFR-1 were detected in vascular smooth muscle (VSM), airway smooth muscle, and airway epithelial cells. FGF-2 was localized in the cytoplasm of airway epithelium and in the nuclei of airway smooth muscle, VSM, and endothelial cells. In COPD cases, an unequivocal increase in FGF-2 expression was observed in VSM (3-fold, P = 0.001) and endothelium (2-fold, P = 0.007) of small pulmonary vessels with a luminal diameter under 200 micro m. In addition, FGFR-1 levels were elevated in the intima (1.5-fold, P = 0.05). VSM cells of large (> 200 micro m) pulmonary vessels showed increased staining for FGF-1 (1.6-fold, P < 0.03) and FGFR-1 (1.4-fold, P < 0.04) in COPD. Pulmonary vascular remodeling, assessed as the ratio of alpha-smooth muscle actin staining and vascular wall area with the lumen diameter, was increased in large vessels of patients with COPD (P = 0.007) and was inversely correlated with FEV(1) values (P < 0.007). Our results suggest an autocrine role of the FGF-FGFR-1 system in the pathogenesis of COPD-associated vascular remodeling.

Arterial Flow Induces Changes in Saphenous Vein EndotheliumProteins Transduced by Cation Channels

Objectives: expression of leukocyte adhesins and proteins controlling thrombosis is likely to be an important determinant of graft patency early following vein bypass. We have previously demonstrated rapid increase in endothelial ICAM-1 and nitric oxide synthase (eNOS) concentrations in human saphenous vein exposed to arterial flow. The aim of this study was to investigate whether ion-channel-blocking drugs could alter these flow-induced changes. Methods: human saphenous vein segments, freshly excised from patients, were placed in a validated in vitro circuit using flow conditions shown to simulate arterial or venous circulations for 90 min, in the presence or absence of ion-channel blockers. The concentrations of ICAM-1, VCAM-1, eNOS and tissue factor (TF) were assessed by quantitative immunohistochemistry in vein exposed to flow and compared with that in freshly excised vein from the same patient. The endothelial protein concentration was calculated as the mean area of staining as percentage of that for the control protein CD31, using computer-aided image analysis. Results: after arterial flow conditions the area ratio of ICAM-1 increased from 21.4±1.4 to 44.6±2.0%, of eNOS increased from 50.0±5.6 to 70.1±5.0%, of VCAM-1 decreased from 16.6±3.4 to 3.6±1.0%, whereas TF staining area ratio was unchanged. Inclusion of the non-selective K+channel blocker, tetraethylammonium, in the arterial perfusion solution abolished all these arterial flow-induced changes. Inclusion of the K+ATP channel blocker, glibenclamide, selectively abolished the arterial flow-induced changes in ICAM-1 and VCAM-1. Inclusion of the calcium channel blocker, nifedipine, abolished the arterial flow-induced changes in eNOS and VCAM-1 but increased the TF staining area ratio from 3.0±0.4 to 8.5±0.7%, p=0.01. Inclusion of the stretch-activated cation-channel blocker, gadolinium, enhanced the arterial flow-induced increase in eNOS, but prevented the arterial flow-induced increase in ICAM-1.Conclusions: perfusion of veins under arterial flow conditions with gadolinium was associated with low endothelial concentrations of ICAM-1, VCAM-1 and TF, but high levels of eNOS. Such a concentration of endothelial proteins may be advantageous in newly implanted vein grafts. In contrast, nifedipine could have adverse effects by promoting increase in TF concentration.

Development of an in vitro model to study the response of saphenous vein endothelium to pulsatile arterial flow and circumferential deformation

To develop an in vitro model of human saphenous vein bypass to facilitate study of the early adaptive responses of venous endothelium to arterial flow conditions. DESIGN MATERIAL AND METHODS: Segments of human saphenous vein (with or without external polytetrafluoroethylene (PTFE) stents to limit circumferential and radial deformation) were mounted in a bypass circuit and subjected to pulsatile flow with oxygenated Krebs solution to simulate arterial or venous flow conditions for a period of 90 min. The viability of the vein was assessed by the tissue ATP concentration and vasomotor responses to phenylephrine, sodium nitroprusside and bradykinin (endothelium-dependent). Immunohistochemistry was used to assess both endothelial preservation (CD31) and the expression of proteins involved in leukocyte adhesion: E-selectin, P-selectin and ICAM-1. Freshly excised veins were used as controls. The concentration of ATP was 320 +/- 11 nmol/g in freshly excised vein (n = 8) and following exposure to the arterial flow circuit increased to 566 +/- 60 nmol/g (n = 8, paired t-test, p = 0.003) in unstented veins and to 421 +/- 49 nmol/g (n = 8, paired t-test, p = 0.002) in externally stented veins (with PTFE). Both endothelium-dependent and sodium nitroprusside-induced vasodilatation responses were preserved after veins were exposed to the arterial flow circuit, but the sensitivity to phenylephrine was increased: EC50 decreasing from 9 microM, p = 0.008. There was a 5-10% decrease in staining area for CD31 after veins, stented or unstented, were exposed to the arterial flow circuit. However, after exposure to the arterial flow circuit, the staining area ratio for ICAM-1/CD31, which remained unchanged in externally stented veins, increased two-fold in unstented veins, p > 0.01: there were no changes in the staining area ratio P-selectin/CD31 and no staining for E-selectin was observed. Vasomotor responses and tissue ATP concentration indicate that the viability of saphenous vein can be maintained for up to 90 min in an ex vivo flow circuit and the CD31 staining indicated endothelial preservation. This opens up the possibility of investigating the early changes in saphenous vein endothelium following exposure to arterial pressure, as at bypass surgery. First results suggest that there is rapid upregulation of the leukocyte adhesion molecule ICAM-1, which can be prevented by limiting the circumferential deformation of the vein with an external PTFE stent.