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Previously developed Asn-Gly-Arg (NGR) peptide-modified multifunctional poly(ethyleneimine)-poly(ethylene glycol) (PEI-PEG)-based nanoparticles (TPIC) have been considered to be promising carriers for the co-delivery of DNA and doxorubicin (DOX). As a continued effort, the aim of the present study was to further evaluate the interaction between TPIC and human umbilical vein endothelial cells (HUVEC) to better understand the cellular entry mechanism. In the present investigation, experiments relevant to co-localization, endocytosis inhibitors and factors influencing the internalization were performed. Without any treatment, there was no co-localization between aminopeptidase N/CD13 (APN/CD13) and caveolin 1 (CAV1). However, co-localization between CD13 and CAV1 was observed when cells were incubated with an anti-CD13 antibody or TPIC. As compared with antibody treatment, TPIC accelerated the speed and enhanced the degree of co-localization. TPIC entered HUVEC not only together with CD13 but also together with CAV1. However, this internalization was not dependent on the enzyme activity of CD13 but could be inhibited by methyl--eyclodextfin (MCD), further identifying the involvement of caveolae-mediated endocytosis (CvME). This conclusion was also verified by endocytosis inhibitor experiments.
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In order to adapt to the training target of medical laboratory technology professionals under the condition of five to four educational system,we reformed the experimental teaching of medical laboratory technology.The experiment teaching was divided three phases (traditional classroom teaching + simulated clinical laboratory training + clinical laboratory teaching).Classroom teaching focused on the cultivation of students' basic skills test;simulation laboratory training focused on strengthening the students' operating skills and basic post ability and professional quality;hospital laboratory teaching focused on strengthening the students' ability of medical laboratory management,test comprehensive analysis and scientific research.The reform practice of two sessions of the students showed that the students' examination skills and comprehensive quality improved significantly,which could better adapt to the social demand for technical personnel.
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Objective To investigate the renoprective effect and its possible mechanism of Niaoduqing on the adriamycin(ADR)-induced nephropathy rats. Methods Forty eight male Sprague-Dawley rats were randomly divided into normal control(n=12), ADR-induced nephropathy(model, n = 12), Benazepril-treated ADR nephropathy(Benazepril, n=12)and Niaoduqing-treated ADR nephropathy(Niaoduqing, n =12)groups. The rat nephropathy model was established by adriamycin injection and unilateral nephrectomy. The rats were sacrifficed per batch at the 4th and 8th weekend.The pathological change of nephridial tissue, the 24-hour urinary protein excretion and renal function were examined. Immunohistochemistry was used to meassure the expression of fibronection(FN), collagenⅣ(COLⅣ), osteopontin(OPN). Results The 24-hour urinary protein excretion, BUN, Scr, triglyeride(TG), cholesterol(Cho)in model group were significantly higher than those in normal group(P<0.01), as well as more server glomerulosclerosis in kidney were observed in model group than those in control group(P<0.01), while the albumin(Alb)was lower (P<0.01). Compared with model group, the 24-hour urinary protein excretion, BUN, Scr, TG, Cho were significantly reduced and renal glomerulosclerosis was improved in Niaoduqing group(P< 0.01), while the Alb was higher(P<0.01). Conclusion Niaoduqing palys an important role in the prevention and treatment for nephropathy.
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Objective To establish a mini pig model suitable for interventional studies in vivo. Methods The endothelia of unilateral renal arteries in 8 purebred Chinese experimental mini pigs(CEMP)was denuded by inflated balloons after the animals were fed with high cholesterol diet for 13 weeks.The CEMP were fed with h high cholesterol diet continuously till the 40th week.The levels of blood lipid panel and creatinine were tested at week 1,14 and 40.Bilateral renal arteries were examined with intravascular uhrasonography at week 14 and 40.The vessel samples were collected at week 40 and stained with haematoxylin-eosin,Masson trichrome technique, oil O and anti-macrophage immunohistological technique. Results Significant differences of blood lipid panel and creatinine were found between week 1 and week 40.Focal ischemic renal injury could be observed pathologically.Renal arteries of CEMP were suitable for interventional procedure such as angiography and intravascular ultrasonography.Cross-sectional information of vessels could be provided clearly by intravascular ultrasonography and the intimamedian thickness of injured renal arteries was much thicker than that of non-injured ones[(0.89±0.03)mm vs (0.30±0.02)mm,P＜0.05]as evidenced by this diagnostic technique.Pathological findings demonstrated the atheroselerotic profiles of the injured renal arteries.Fibrous and fibro-fatty plaques were the main pathologic types in this CEMP model. Conclusions An animal model with renal arterial atherosclerosis mimicking the progression of atheroselerotic renovaseular disease,which is suitable for interventional procedure is established successfully.Intravascular ultasonography may have potential clinical prospect on the evaluation of atherosclerotic renovaseular disease.
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OBJECTIVE@#To study the expressions of Cyclin D1 and p16 proteins in hypopharyngeal squamous cell carcinoma and their clinical significance.@*METHOD@#Immunohistochemical technology (P-V) was applied to detect the expression of Cyclin D1 and p16 in 36 cases of hypopharyngeal squamous cell carcinoma and 10 cases of normal epithelium.@*RESULT@#(1) The expression of cyclin D1 in the tumorous cell was significantly higher than that in normal epithelium (P 0.05); (3) There was correlation between the expression of Cyclin D1 and the expression of p16 (r(s) = -0.420, P < 0.05).@*CONCLUSION@#The over-expression of Cyclin D1 and the under-expression of p16 may play a significant role in the occurrence incidence and development of hypopharyngeal squamous cell carcinoma, and may be important indicators for cervical lymph node metastases.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Metabolism , Pathology , Case-Control Studies , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase Inhibitor p16 , Metabolism , Hypopharyngeal Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , PrognosisABSTRACT
Objective To investigate the feasibility of gene modification of peripheral blood derived endothelial progenitor cells(EPCs).Methods The rabbit mononuclear cells were collected from rabbit peripheral blood and cultured.The uptaking test of DiI-Ac-LDL was directly visualized with fluorescent microscopy,the immunofluorescence analysis was performed to detect the expression of surface marker.And the human tissue kallikrein(HTK)gene was amplified with PCR,and inserted into pEGFP-N3 vector.EPCs were transfected with the constructed plasmid by means of lipidosome,and the HTK-EGFP fused protein was visualized directly with fluorescent microscopy,and the expression of HTK was detected by RT-PCR and ELISA.Results After 5～6 days of culture,spindle-shaped attached cells clustered together,and cobblestone-like cell layer appeared after 15 days.Cells were positive of uptaking DiI-Ac-LDL and expressing vWF and CD133 related antigen.Plasmids were correctly formed and expressed in the rabbit EPCs.The fused protein had activity of both HTK and EGFP.Conclusion The HTK gene modified EPCs is potential in the treatment of artery injury and endothelial dysfunction.