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Objective: To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinase-2 (TIMP-2) and on cell migration of cultured rat vascular smooth muscle cells (VSMCs). Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods: Rat VSMCs were incubated with different concentrations of homocysteine (50–5 000 μmol/L). Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin (10−9–10−5 mol/L) were added when VSMCs were induced with 1 000 μmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results: Homocysteine (50–1 000 μmol/L) increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5 000 μmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and activation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine (50–5 000 μmol/L) stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions: Homocysteine (50–1 000 μmol/L) significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression and activation of MMP-2 and abnormal migration of VSMCs induced by homocysteine.  相似文献   

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目的:研究离体热处理和顺铂联合热处理对术中回收血红细胞功能的影响及其中混杂的肝肿瘤细胞株(HepG2)的杀灭作用。创新点:(1)采用多种评价指标研究了不同时间离体热处理对术中回收血中混杂的HepG2的杀灭作用及对红细胞的影响,并确定了对红细胞安全且能有效杀灭HepG2的离体热处理时间。(2)从多个角度评价了离体顺铂联合热处理对术中回收血红细胞的影响及对其中混杂的HepG2的杀灭作用,确定了该方案中对红细胞安全且能有效去除HepG2的顺铂浓度。方法:采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)、5-乙炔基-2’脱氧尿嘧啶核苷(EdU)和平板克隆形成评估HepG2的细胞存活率、DNA复制率和克隆形成能力(图1和3);从红细胞渗透脆性、携氧能力(2,3-二磷酸甘油酸(2,3-DPG)、半饱和氧分压(P50))、能量代谢(Na+-K+-ATPase、pH)、膜完整性(游离血红蛋白(Hb)、血清K+和Na+浓度、细胞膜磷脂酰丝氨酸外翻比例)等角度评估红细胞功能(图2和4;表3)。结论:肝肿瘤术中回收血经离体顺铂联合热处理(42°C,50μg/ml)60min后,能有效清除其中混杂的HepG2,但对红细胞无显著影响,值得体内进一步研究顺铂热处理有效应用于肿瘤手术自体血液回输的安全方案。  相似文献   

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To conduct a systematic review of group studies assessing the association of serum vitamin D status with the severity of liver fibrosis in chronic hepatitis C patients using meta-analysis. The relevant research literatures were identified by searching PubMed and EMBASE databases prior to October 2013 with no restrictions. We included group studies that reported odds ratio (OR) estimates with 95% confidence intervals (CIs) or a mean with standard deviation (SD) for the association between serum vitamin D status and the severity of liver fibrosis in chronic hepatitis C patients. Approximately 8321 participants from several countries were included in this analysis. Six studies on serum vitamin D status and the severity of liver fibrosis were included in this meta-analysis. ORs with 95% CIs were extracted from four studies and the pooled ORs were 0.866 (95% CI, 0.649 to 1.157). The means with SDs were extracted from three studies and the pooled means were −0.487 (95% CI, −0.659 to −0.315). There was statistically significant heterogeneity among the mean data extracted studies (P=0.029; I 2=71.8%) but not among the OR data extracted studies (P=0.061; I 2=55.6%). Finally, results from the mean data extracted studies suggest that lower serum vitamin D is a risk factor for the severity of liver fibrosis in chronic hepatitis C patients. However, there is no conclusive evidence on this association because of inconsistencies between the OR data extracted studies and the mean data extracted studies.  相似文献   

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β-glucanase was purified from a solid-state culture ofTrichoderma reesei on wheat bran in three steps which comprised ammonium sulfate precipitation, Sephadex G-100 chromatography, and DEAE-Sephadex A-50 chromatography. The molecular mass was determined to be 35.21 kilodaltons by sodium dodecyl sulfate-12.5% polyacrylamide gel electrophoresis. The β-glucanase at low pHs was more stable than that at high pHs, and optimum pH was 5.0. The optimum temperature was 60°C, and β-glucanase was relatively stable at below 40°C for 60 min. TheK m of the enzyme on β-glucan was 10.86 mg/ml, and theV max on β-glucan was 14286 μmol of glucose equivalents per mg of the pure enzyme per min. The β-glucanase activity was significantly inhibited by Fe3+ ions, and was reduced in the presence of Cu2+ ions, Mn2+ ions and Mg2+ ions at 5 mmol/L and 10 mmol/L, respectively. The β-glucanase activity was stimulated by Co2+ ions, Ca2+ ions, Zn2+ ions, and Fe2+ ions at 1 mmol/L and 5 mmol/L, respectively. Project supported by Foundation for University Key Teacher of the State Ministry of Education, the National Natural Science Foundation of China (No. 30000118) and Zhejiang Provincial Natural Science Foundation of China (No. 399409).  相似文献   

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1 Introduction Ahugeamountofliteratureexistsonthestudyofi dentifyingtracemetalionsbymeansofspectropho tometry[1 3] ,fluorimetry[4 6 ] aswellas polarogra phy[7 9] ,whichhaveactiononlocatingtheendpointoftheindicatingreaction .Howeverbothspectropho tometryandfluorimetryareeasytobeinterferedbycolororprecipitation ,whilepolarographyhasanissueofmercurypollution .Thecatalyzingkineticanalysishasthecharacteristicsofhighsensitivityandspeedi ness.Oscillographicpotentiometryisasimpleintu itionisticanal…  相似文献   

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Liu  Yongling  Wu  Songhai  Jia  Shaoyi  Liang  Zongsheng  Han  Xu 《天津大学学报(英文版)》2019,25(4):371-380
Natural pyrites contain high levels of adsorbed and structurally incorporated arsenic(As),which may simultaneously result in the release of As and affect the oxidation process of pyrite.However,the oxidation and electrochemical behaviors of As on the oxidation reactivity of pyrites are still not clear.In this study,pyrite was prepared by a hydrothermal method and applied to study the oxidation mechanism between pyrite and aqueous arsenate.Analyses of X-ray diffraction,X-ray photoelectron spectroscopy,and scanning electron microscopy demonstrate that the as-prepared sample is an octahedron-like pyrite with high purity and crystallinity.The interaction between As(V)and pyrite as well as the electrochemical behaviors of pyrite oxidation in the presence of aqueous arsenate were investigated under acidic conditions by an ion analysis method,cyclic voltammetry(CV),Tafel,and electrochemical impedance spectroscopy(EIS).The results of the chemical reaction indicate that electrons are transferred from S _2~(2-)to dissolved oxygen with the formation of SO _4~(2-)in the initial As(V)concentration range of 0–0.3 mmol/L.In the initial As(V)concentration range of 0.4–1.2 mmol/L,electrons are transferred from S _2~(2-)to As(V)with the formation of elemental S ~(0 )and As(III).The CV,the Tafel plot and EIS analyses indicate that aqueous arsenate in an electrolyte promotes oxidation reactivity and passivation of the pyrite electrode.Moreover,the electron transfer rate increases with increasing aqueous arsenate concentration in the electrolyte.  相似文献   

9.

Objective

The efflux pump (EP) is one of the major mechanisms of antibiotic resistance in Klebsiella pneumoniae. However, there are few reports on the effect of the abuse of antibiotic use on the activity of EPs. To determine whether the use of low efficacy antibiotics has any effect on the activity of EPs and induces drug resistance in K. pneumoniae, we investigated the effect of ciprofloxacin on the activity of EPs in K. pneumoniae strains.

Methods

Sixteen susceptible K. pneumoniae strains were isolated from patients and their minimum inhibitory concentrations (MICs) of ciprofloxacin were measured in the absence and presence of the pump inhibitor carbonyl cyanide m-chlorophenyl hydrazone (CCCP). The strains were then induced with a gradient of ciprofloxacin until the MICs of the strains showed no further increase, to obtain induced resistant strains. The EP activities of the strains before and after induction were compared using EP inhibition and ethidium bromide (EtBr) accumulation assays.

Results

The MIC values of the strains were 16–256 times higher after induction than before induction. In the presence of CCCP, the MIC values of 50% of the induced strains were 2–4-fold lower than that in the absence of this inhibitor. The EtBr accumulation assay showed that the fluorescence of EtBr in the induced cells was lower than that in the cells before induction.

Conclusions

EPs are widespread in susceptible and drug-resistant K. pneumoniae strains. Induction with ciprofloxacin may increase the activity of EPs in K. pneumoniae. The EtBr accumulation assay is more sensitive than the EP inhibition assay in evaluating the activity of EPs in K. pneumoniae.  相似文献   

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The objective was to investigate the expression of bone morphogenetic protein (BMP) family members in the mouse uterus during the estrous cycle by real-time polymerase chain reaction (PCR) and immunohistochemistry. Uterine samples from Swiss ICR mice were collected and dissected free of surrounding tissue. One uterine horn was snap frozen in liquid nitrogen immediately after collection and stored at −80 °C for RNA extraction, and the other was fixed in 40 mg/ml paraformaldehyde at room temperature for immunolocalization of BMP2 protein. Real-time PCR analysis showed that the expression level of Bmp2 was significantly higher at proestrus than at estrus and metestrus (P<0.05). The relative abundance of Bmp4 exhibited significant fluctuations, but there were no statistically significant differences between the expression levels of Bmp2 and Bmp4 (P>0.05). The expression levels of Bmpr1a and Bmpr2 remained unchanged during estrous cycles. However, the level of Bmpr1b mRNA decreased significantly at estrus (P<0.05), increasing subsequently at metestrus. Furthermore, the level of Bmpr1b mRNA was significantly lower than those of Bmpr1a and Bmpr2 mRNA at the corresponding stages (P<0.05). All three receptor-regulated Smads (R-Smads) detected were differentially expressed in the mouse uterus and the expression levels of Smad1 and Smad5 were significantly higher than that of Smad8 (P<0.05). In addition, the expression level of Smad4 did not change substantially throughout the estrous cycle. Immunohistochemical experiments revealed that BMP2 protein was differentially expressed and localized mainly in the uterine luminal and glandular epithelial cells throughout the estrous cycle. In conclusion, our results provide information about the variation in the mRNA levels of Bmp2 and Bmp4 and related components of the BMP signaling pathway. The data provide quantitative and useful information about the roles of endometrial BMP proposed and demonstrated by others, such as the degradation and remodeling of the endometrium.  相似文献   

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研究目的:优化获得血红密孔菌(P.sanguineus)的最佳培养基组成,提高耐热漆酶的产量。创新要点:获得了目前文献报道的最高水平的漆酶活力。研究方法:通过单因素试验研究了不同培养基(番茄汁、麦麸、麦芽提取物和葡萄糖细菌蛋白胨培养基)和不同组合诱导剂(大豆油、阿魏酸、没食子酸、二甲基苯胺、酸性蓝62和活性蓝19分别与硫酸铜组合诱导剂)对P.sanguineus产耐热漆酶的影响。在此基础上采用中心组合试验设计,进一步研究了番茄汁培养基结合硫酸铜和大豆油组合诱导剂对P.sanguineus产耐热漆酶的影响。利用SAS10.0和响应面分析方法对试验结果进行了统计分析和建立回归模型。重要结论:通过中心组合设计优化得出P.sanguineus产耐热漆酶的最优培养基条件:以36.8%番茄汁为培养基,以3 mmol/L硫酸铜和1%大豆油作为组合诱导剂。该条件下在10 L搅拌槽生物反应器中漆酶活力达到了143 000 IU/L(2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)为底物,pH值为3.0)。  相似文献   

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The dense granule protein 4 (GRA4) is a granular protein from Toxoplasma gondii, and is a candidate for vaccination against this parasite. In this study, the plasmid pcDNA3.1-GRA4 (pGRA4), encoding for the GRA4 antigen, was incorporated by the dehydration-rehydration method into liposomes composed of 16 mmol/L egg phosphatidylcholine (PC), 8 mmol/L dioleoyl phosphatidylethanolamine (DOPE), and 4 mmol/L 1,2-diodeoyl-3-(trimethylammonium) propane (DOTAP). C57BL/6 mice and BALB/c mice were immunized intramuscularly three times with liposome-encapsulated pGRA4 to determine whether DNA immunization could elicit a protective immune response to T. gondii. Enzyme-linked immunosorbent assay (ELISA) of sera from immunized mice showed that liposome-encapsulated pGRA4 generated high levels of IgG antibodies to GRA4. Production of primary interferon (IFN)-γ and interleukin (IL)-2 in GRA4-stimulated splenocytes from vaccinated mice suggested a modulated Th1-type response. 72.7% of C57BL/6 mice immunized with liposome-encapsulated pGRA4 survived the challenge with 80 tissue cysts of ME49 strain, whereas C57BL/6 mice immunized with pGRA4 had only a survival rate of 54.5%. When immunized BALB/c mice were intraperitoneally challenged with 103 tachyzoites of the highly virulent RH strain, the survival time of mice immunized with liposome-encapsulated pGRA4 was markedly longer than that of other groups. Our observations show that liposome-encapsulated pGRA4 enhanced the protective effect against infection of T. gondii. Project supported by the Science Foundation of the Health Bureau of Zhejiang Province, China (Nos. 2003QN003 and 2005A001) and the Science Foundation of the Science and Technology Department of Zhejiang Province, China (No. 2006C13022)  相似文献   

15.
Transgenic Brassica compestris L.spp.chinensis plants expressing a choline oxidase(codA) gene from Arthrobacter globiformis were obtained through Agrobacterium tumefaciens-mediated transformation.In the transgenic plants,codA gene expression and its product transportation to chloroplasts were detected by the enzyme-linked immunosorbent assay(ELISA) examination,immunogold localization,and 1 H-nuclear magnetic resonance( 1 H-NMR) . Stress tolerance was evaluated in the T3 plants under extreme temperature and salinity conditions.The plants of transgenic line 1(L1) showed significantly higher net photosynthetic rate(Pn) and Pn recovery rate under high(45°C,4 h) and low temperature(1°C,48 h) treatments,and higher photosynthetic rate under high salinity conditions(100,200,and 300 mmol/L NaCl,respectively) than the wild-type plants.The enhanced tolerance to high temperature and high salinity stresses in transgenic plants is associated with the accumulation of betaine,which is not found in the wild-type plants.Our results indicate that the introduction of codA gene from Arthrobacter globiformis into Brassica compestris L.spp.chinensis could be a potential strategy for improving the plant tolerance to multiple stresses.  相似文献   

16.
Objective: To evaluate the immunological effects of three types of domestic 10-μg/dose hepatitis B vaccines in adults compared with a foreign vaccine, and to provide scientific evidence in support of adult hepatitis B vaccination. Methods: Adults from five counties (Deqing, Changxing, Nanxun, Wuxing, Anji) in Huzhou City, Shaoxing County and Tongxiang County, Zhejiang Province, China were selected. Blood samples were taken to assess serum HBsAg, anti-HBs, and anti-HBc using a chemiluminescence immunoassay. Adults, aged 16 to 49 years and who were anti-HBs negative at baseline, received hepatitis B immunizations at 0, 1, and 6 months. Anti-HBs levels were assessed one month after the third and final vaccination. Results: A total of 1 872 adults were immunized and the average positive rate was 89.5%. Four types of hepatitis B vaccine were used, including three from Chinese companies (Shenzhen Kangtai, Dalian High-Tech, and North China Pharmaceutical) and one from a UK company (GlaxoSmithKline). Their seroconversion rates were 81.67%, 95.05%, 89.64%, and 86.81%, respectively. There was a significant difference between the anti-HBs positive conversion rates of the four types (P<0.005) but the seroconversion rates among the different vaccines were not significantly different (χ 2=2.123, P=0.145). The average anti-HBs geometric mean titers (GMTs) of non-immune adults immunized with each of the four vaccines were 177.28, 473.23, 246.13, and 332.20 mIU/ml, respectively. There were no statistically significant differences in the GMTs between the three types of domestic vaccine and the foreign vaccine (t=−1.575, P=0.116). Conclusions: Domestic recombinant hepatitis B vaccines can achieve immunization effects comparable to those of a foreign vaccine.  相似文献   

17.

Objectives

To investigate the effects of different nitrate sources on the uptake, transport, and distribution of molybdenum (Mo) between two oilseed rape (Brassica napus L.) cultivars, L0917 and ZS11.

Methods

A hydroponic culture experiment was conducted with four nitrate/ammonium (NO3 ?:NH4 +) ratios (14:1, 9:6, 7.5:7.5, and 1:14) at a constant nitrogen concentration of 15 mmol/L. We examined Mo concentrations in roots, shoots, xylem and phloem sap, and subcellular fractions of leaves to contrast Mo uptake, transport, and subcellular distribution between ZS11 and L0917.

Results

Both the cultivars showed maximum biomass and Mo accumulation at the 7.5:7.5 ratio of NO3 ?:NH4 + while those were decreased by the 14:1 and 1:14 treatments. However, the percentages of root Mo (14.8% and 15.0% for L0917 and ZS11, respectively) were low under the 7.5:7.5 treatment, suggesting that the equal NO3 ?:NH4 + ratio promoted Mo transportation from root to shoot. The xylem sap Mo concentration and phloem sap Mo accumulation of L0917 were lower than those of ZS11 under the 1:14 treatment, which suggests that higher NO3 ?:NH4 + ratio was more beneficial for L0917. On the contrary, a lower NO3 ?:NH4 + ratio was more beneficial for ZS11 to transport and remobilize Mo. Furthermore, the Mo concentrations of both the cultivars’ leaf organelles were increased but the Mo accumulations of the cell wall and soluble fraction were reduced significantly under the 14:1 treatment, meaning that more Mo was accumulated in organelles under the highest NO3 ?:NH4 + ratio.

Conclusions

This investigation demonstrated that the capacities of Mo absorption, transportation and subcellular distribution play an important role in genotype-dependent differences in Mo accumulation under low or high NO3 ?:NH4 + ratio conditions.
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18.
A highly sensitive amperometric sulfadiazine sensor fabricated by electrochemical deposition of poly(cobalt tetraaminophthalocyanine) (poly(CoIITAPc)) on the surface of a multi-walled carbon nanotubes-Nafion (MWCNTs-Nafion) modified electrode is described. This electrode showed a very attractive performance by combining the advantages of CoIITAPc, MWCNTs, and Nafion. Compared with the bare glassy carbon electrode (GCE) and the MWCNTs-Nafion modified electrode, the electrocatalytic activity of poly(CoIITAPc)-coated MWCNTs-Nafion GCE generated greatly improved electrochemical detections toward sulfadiazine including low oxidation potential, high current responses, and good anti-fouling performance. The oxidation peak currents of sulfadiazine obtained on the new modified electrode increased linearly while increasing the concentration of sulfadiazine from 0.5 to 43.5 μmol/L with the detection limit of 0.17 μmol/L.  相似文献   

19.
The effects of salinity(50 mmol/L NaCl) and Cd(1 μmol/L CdCl2) as sole and combined on growth and photosyn-thetic parameters were studied using two soybean genotypes,Huachun 18 and NGB.The concentrations of Cd2 ,Zn2 ,Ca2 ,Mg2 ,K and Na were also determined in seeds and pods.Huachun 18 suffered a more serious decrease than NGB in net photosynthetic rate(Pn) in the treatments of salinity stress alone and combined stress(NaCl Cd),showing that it is relatively sensitive to salinity.The decrease in Pn caused by salt stress in Huachun 18 was mainly due to the reduced total chlorophyll content and photosynthetic efficiency(the ratio of variable fluorescence to maximal fluorescence,Fv/Fm),whereas the decease in NGB was mainly related to reduced stomatal conductance(Gs).The combined stress of both Na and Cd did not induce further decrease in photosynthesis and fluorescence in the two genotypes relative to salt or Cd stress alone.Greater change in the pod concentrations of Zn2 ,Ca2 ,Mg2 ,K and Na was detected under salt stress for Huachun 18 than for NGB.The results suggested that the interactive effect of NaCl-Cd on growth and nutrient uptake differs between the two soybean genotypes.  相似文献   

20.
Zeng  Qingyou  Jia  Shaoyi  Gong  Yufeng  Wu  Songhai  Han  Xu 《天津大学学报(英文版)》2019,25(6):567-575

Cr(VI) and phenol are toxic contaminants that need to be treated, and different methods have been researched to simultaneously remove these two contaminants from industrial wastewater. In this study, Cr(VI) was used as a novel Fenton-like catalyst in phenol degradation by H2O2. In the pH range of 3.0‒11.0, the degradation efficiency of phenol decreased with elevated pH. At pH = 3.0, 100 mg/L phenol was effectively degraded by 2 mmol/L Cr(VI) and 20 mmol/L H2O2. At pH = 7.0 and the same conditions as those of pH = 3.0, 79% of 100 mg/L phenol was removed within 6 h, which was an improvement in pH limitation compared with the Fe(II)-mediated Fenton reaction. Quenching experiments indicated that ·OH generated from the catalysis of H2O2 by Cr(V) instead of Cr(VI) was the primary oxidant that degraded phenol. When pyrophosphate was added in the Cr(VI)/H2O2 system, complexes with the Cr(V) intermediate rapidly formed and inhibited H2O2 decomposition, implying that the decomposition of H2O2 to ·OH was catalyzed by Cr(V) instead of Cr(VI). The presence of anions such as chloride and sulfate had insignificant effect on the degradation of phenol. TOC and UV analyses suggest that phenol could not be completely oxidized to CO2 and H2O, and the intermediates identified by high performance liquid chromatography further indicates that maleic acid and benzoquinone were intermediates which may be further degraded into short chain acids, primarily maleic, formic, acetic, and oxalic acids, and eventually into CO2 and H2O. Considering that more than 50% Cr(VI) can also be removed during this process, the Cr(VI)/H2O2 system is more appropriate for the simultaneous removal of Cr(VI) and phenol contaminants from industrial wastewater.

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