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Objective:To construct tissue engineering small-caliber anti-calcifiction blood vessels with micron slow-release magnesium chloride.Methods:After decellularizing sheep carotid artery by combining Triton X-100+ deoxycholate sodium salt and DNA/RNA ribozyme, tissue engineering small-caliber vascular scaffold was made, HE staining of elastic fiber and collagen were carried out at the same time, and scanning electron microscope was used to observe the decellularization and the performance of vascular stent. The microemulsion anti-calcification slow-release microsphere particles loaded with magnesium chloride(MgCl 2) were prepared by double emulsion method, ultrasonic breaking, high speed stirring and evaporation method. Detected the particle size, encapsulation rate, drug loading(rate) of the sustained-release microspheres and measured the sustained-release curve. After the artificial small-caliber blood vessel was cross-linked with carbodiimide hydrochloride/succinic imine(EDC/NHS), freeze-drying technology was used to combine the micron slow-release microspheres loaded with MgCl 2 with the vascular scaffold. Observed the combination under the electron microscope, and tested the thickness and tensile strength of the specimen blood vessels. Results:After decellularization, the sheep carotid artery could remove all kinds of cells and maintain the original performance of the scaffold. The averaged particle size of micro-calcium-resistant slow-release microspheres loaded with MgCl 2 was(1.31±0.02)μm, which was relatively uniform. The encapsulation rate of microsphere particles was 82.79%, and the drug loading(rate) was 2.98%, which existed within 25 days slow release, the release rate reached 81.08%. The slow-release microsphere particles loaded with chlorinase could be effectively and tightly combined with small-caliber tissue engineering blood vessels. Conclusion:The slow-release microsphere particles loaded with magnesium chloride made of PLGA as a carrier have the effect of slow-release magnesium ions. It laid the foundation for the construction of anti-calcification tissue engineering small-caliber blood vessels.
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Objective To investigate the role of autophagy in high glucose-induced podocyte lipid droplet metabolism.Methods (1) Cultured,conditionally immortalized human podocytes (HPC)were divided into normal control group,high glucose group and mannitol group.Oil red O staining and oil red O staining extraction assay was used to observe the degree of lipid accumulation;Protein level of SREBP-1 was analyzed by Western blotting.(2) HPC were cultured and divided into normal control group,high glucose group,high glucose+3-methyladenine (3-MA) group,and mannitol group.Acridine orange staining was used to observe the formation of autophagosomes.Western blotting was used to detect the protein levels of beclin-1 and LC3-Ⅱ/LC3-Ⅰ.Oil red O staining and oil red O staining extraction assay was used to observe the degree of lipid accumulation;Western blotting was used to analyze the expression of SREBP-1.Results (1) Compared with the normal control group,the lipid accumulation in the high glucose group was increased and the lipid metabolism related molecule SREBP-1 was up-regulated (P ＜ 0.05);There was no significant difference between the normal control group and the mannitol group in lipid accumulation (P ＞ 0.05).(2) Compared with the normal group,the number of autophagosomes was increased and autophagy-related proteins beclin-1 and LC3-Ⅱ/LC3-Ⅰ were up-regulated in high glucose group (all P ＜ 0.05).After intervened with 3-methyladenine,a significant decrease in autophagosomes was observed;Protein levels of autophagy-related proteinsbeclin-1 and LC3-Ⅱ/LC3-Ⅰ were decreased (all P ＜ 0.05);The lipid droplets in the high glucose+3-MA group was decreased and lipid metabolism related molecule SREBP-1 was down-regulated (all P ＜0.05).Conclusion Autophagy may be involved in the process of high-glucose-induced podocyte lipid accumulation by affecting SREBP-1 expression,and inhibition of autophagy can alleviate the high-glucose-induced podocyte lipid accumulation.
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Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of ATP-binding cassette transporter A1(ABCA1) in Ang Ⅱ-infused rat model and cultured human podocytes,and to explore the role of ABCA1 in Ang Ⅱ-induced cholesterol accumulation of podocytes.Methods Twelve Wistar rats were randomly subjected to normal saline infusion,or Ang Ⅱ infusion at 400 ng· kg-1 · min-1 (via subcutaneous osmotic minipumps) for 8 weeks.The expression of glomerular ABCA1 was analyzed by Western blotting and real-time fluorescent quantitative PCR.In vitro,conditionally immortalized human podocytes were divided into normal group,Ang Ⅱ group,Ang Ⅱ + scrambled siRNA group,Ang Ⅱ + ABCA1 siRNA group.The expression of podocyte ABCA1 was assessed by immunofluorescence,Western blotting and real-time fluorescent quantitative PCR.Oil Red 0 staining was used to observe the lipid droplets in podocytes and cholesterol efflux assay kit was used to measure the cholesterol efflux rate of podocytes.Fluorescein isothiocyanate (FITC)-conjugated phalloidin was used to observe the podocyte cytoskeleton.Results In vivo,compared with normal group,the protein and mRNA expression of glomerular ABCA1 in Ang Ⅱ-infused rats were decreased (P ＜ 0.05).In vitro,ABCA1 was distributed in the cytomembrane of podocytes,and compared with normal group,Ang Ⅱ treatment down-regulated the expression of ABCA1 (P ＜ 0.05).Increased lipid droplets,decreased cholesterol efflux and cytoskeletal rearrangement were observed in Ang Ⅱ-treated podocytes.When compared to Ang Ⅱ group,podocytes stimulated by Ang Ⅱ and then transfected with ABCA1 siRNA had lower expression level of ABCA1 mRNA and protein (all P ＜ 0.05).More lipid droplets and lower cholesterol efflux rate could be observed in Ang Ⅱ +ABCA1 siRNA group (P＜ 0.05).Conclusion The reduced expression of ABCA1 may be involved in Ang Ⅱ-induced cholesterol accumulation in podocytes.
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Objective To investigate the role of IQ domain GTPase-activating protein 1 (IQGAP1) in angiotensin Ⅱ (Ang Ⅱ)-induced podocyte apoptosis and the underlying mechanism.Methods Differentiated mouse podocytes were exposed to Ang Ⅱ at different concentrations for 6 h or at 10-8 mol/L for variable incubation time.Podocyte apoptosis was assessed by flow cytometry.Expression of IQGAP1 was analyzed by immunofluorescence and Western blotting.IQGAP1 siRNA and MAPK pathway inhibitors(10 μmol/L SB202190,25 μmol/L SP600125,10 μmol/L U0126) were further introduced to investigate the role of IQGAP1 and MAPK signalings in the process.And coimmunoprecipitation was used to evaluate the interaction between ERK1/2 and IQGAP1.Results (1)Ang]] promoted podocyte apoptosis in a dose-and time-dependent manner.(2) IQGAP1 was located in celluar membrane and cytoplasm of cultured podocytes.Exposure to Ang Ⅱ stimulated IQGAP1expression in a dose-and time-dependent manner,and elevated phosphorylation of p38,JNK,and ERK1/2 simultaneously.(3) Pretreatment with SB202190,SP600125,or U0126 dramatically prevented Ang Ⅱ-promoted podocyte apoptosis respectively (P ＜ 0.05).However,the protein level of IQGAP1 was not altered.(4) Knockdown of IQGAP1 with siRNA obviously prevented Ang]Ⅱ-induced apoptosis of podocytes(P ＜ 0.05) and reduced Ang Ⅱ-induced phosphorylation of ERK1/2(P ＜ 0.05),but not that of p38,JNK.This was accompanied by a reduced interaction between ERK1/2 and IQGAP1(P ＜ 0.05).Conclusion IQGAP1 contributes to Ang Ⅱ-induced podocyte apoptosis by interacting with the ERK1/2 signaling protein.
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Objective To identify the differentially expressed microRNAs in the early transformed cells,the late transformed cells and their parental BEP2D cells.Methods The differentially expressed microRNAs in the above cells were identified by microRNAs microarray assay.Results There were 38differentially expressed microRNAs in R15H20 cells versus BEP2D cells,with 18 upregulated and 20downregulated microRNAs.R15H20 and RHT35 cells shared 25 differentially expressed microRNAs compared with BEP2D cells,with 15 down-regulated and 10 up-regulated microRNAs.There were 87differentially expressed microRNAs in RHT35 cells versus BEP2D cells,with 47 upregulated and 40 downregulated microRNAs.There were 38 differentially expressed microRNAs in RHT35 cells versus R15H20 cells with 20 upregulated and 18 downregulated microRNAs.Conclusions microRNAs are differentially expressed in the different stages of carcinogenesis of BEP2D cells induced by α particles,which suggests that microRNAs may play an important role in α particle-induced malignant transformation of BEP2D cells.
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A molecularly imprinted polymer (MIP) using benzoic acid as template molecule, 4-vinyl pyridine (4-VP) as functional monomer, ethylene dimethacrylate (EDMA) as cross-linker, was prepared by bulk polymerization. The needle-type gas concentrator was developed using the MIP as adsorption medium. The device was coupled with gas chromatography (GC) for the analysis of volatile organic compounds (VOCs). The effect of polymerization conditions on adsorption property, such as polymerization time, ratio of the reagents, pre-polymerization time, type of solvents, type of template molecules, has been evaluated. The results of gas chromatographic analysis demonstrated that the optimum conditions for getting the best adsorption performance of the synthesized were polymerization time 6 h at 60 ℃, the ratio of the reagents (template molecule : functional monomer : cross-linker) 1∶ 4∶ 20, pre-polymerization time of 3 h, acetonitrile as solvent, benzoic acid as template.
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A magnetic separator was used to separate magnetic bacteria based on their magnetotactic characteristics. Acidithiobacillus ferrooxidans, a bacterium that could synthesize intra-cellular nanometer magnetic particles, was investigated as an example. Strong magnetic and weak magnetic cells were separated and collected. On average, the number of the magnetic particles present in the strong magnetic cells is more than that of the weak magnetic cells. Moreover, semisolid-plate magnetophoresis showed that the magnetotaxis of strong magnetic cells was stronger than the weak magnetic cells. These results suggest that the magnetic separator can be used to isolate the magnetic bacteria, which will facilitate the research of magnetic bacteria.
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Acidithiobacillus , Metabolism , Bacteria , Metabolism , Bacterial Physiological Phenomena , Bacteriological Techniques , Methods , MagneticsABSTRACT
Coronary stent can cause mechanical injury to tunica intima and stimulation to vessel wall, resulting to platelet and inflammatory cell aggregation and infiltration, release of inflammation mediators, chemotatic factor, adhesion molecule and growth factor, and promoting migration and proliferation of vascular smooth muscle cells. The inflammatory reaction post stenting is highly correlated with intravascular restenosis. The drug-eluting stent against proliferation of vascular smooth muscle cells and inflammatory reaction can reduce the intensity and duration of body inflammatory reaction, improve stent technique, relieve damage of stenting to vessel wall, and reduce incidence of intravascular restenosis.
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Resulting from the shape stability, metal vascular stent has limited the vascular retraction, and subsequently prevent the unfavorable vascular remodeling. However, the metal stent requires further anticoagulant therapy after implantation, induces the hyperplasy of vascular smooth muscle cells, and cannot completely prevent the occurrence of in-stent restenosis. Surface modification of metal stent may reduce thrombogenesis. Based on the metal stent, drug eluting stent can transfer the active drugs to the damaged vessels, release them into the vascular wall and inhibit the in-stent restenosis. The eluting drugs restrain both the proliferation of smooth muscle cells and the regeneration of normal endothelial cells, leading to delay the vascular endothelialization and increase the risk of delayed thrombogenesis. The effect of stent implantation on the modus and size of vascular injury varies according to different operational techniques, processing technologies and designs, thus influencing the occurrence of in-stent restenosis. It is a potential study topic of interventional therapy to develop new eluting materials and eluting drugs, modify formulation, as well as facilitate the stent structure.
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Objectives To evaluate the application of flow-through hybrization and gene chip (HybriMax) on human papillomavious (HPV) subtype detection in clinical.Method 3 086 sexually active females from out-patient department were selected for HPV subtype analysis using HybriMax. Cervical tissue samples were taken under the colposcope from 463 females who had cervical lesion for pathological analysis. The predictive value of HybriMax in cervical abnormality was compared with pathological results, which were used as a golden standard. DNA sequence in HPV E6/E7 region was performed among 80 females with HPV 16 positive by HybriMax to determine the accuracy of HybriMax.Results All 21 different subtypes were found and total HPV positive rate was 63.1%(1 947/3 086). Among the 21 HPV subtypes, 5 of them had a infection rate over 5% and they were HPH16(15.9%, 490/3 086), 58(11.2%, 346/3 086), 52(8.5%、261/3 086)、33(6.3%、195/3 086)、53(6.2%、192/3 086)、6(5.6%、172/3 086), and CP8304(5.0%、155/3 086). The sensitivity for high degree squamous intraepithelial dysplasia(HSIL, CINⅡ+CINⅢ) by HybriMax was 95.49%(95%CI=95.44%～98.27%), while specificity was 34.85%(95% CI= 32.59%～37.57%)、Positive predictive value for HSIL was 37.13%(95%CI=30.79%～40.96%),while negative predictive vale was 95.04%(95%CI=89.24%～98.44%). Eighty sequence results in E6/E7 region completely matched to HybriMax results.Conclusion HybriMax has a high accurate rate in HPV subtype diction with good sensitivity and specificity for HSIL and above. It is an effective method to detect HPV subtype in clinical.
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<p><b>OBJECTIVE</b>To investigate the possible effects of long-term exposure to dust containing thorium and thoron progeny on dust-exposed miners.</p><p><b>METHODS</b>A negative, high voltage, exhaled thoron progeny measurement system was used to estimate the miners' thorium lung burden.</p><p><b>RESULTS</b>The highest thorium lung burden of 638 miners was 11.11 Bq. The incidence of stage 0(+) pneumoconiosis was higher among dust-exposed miners. Lung cancer mortality of the dust-exposed miners was significantly higher than that of controls (P < 0.005).</p><p><b>CONCLUSION</b>There is a difference in cancer rates between those who have long-term exposure to dust containing thorium (in which carcinogenic ThO(2) and SiO(2) exist) and thoron progeny and those who have not.</p>