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Chen B.-D.,Clinical Medical Research Institute | He C.-H.,Clinical Medical Research Institute | He C.-H.,Xinjiang Medical University | Chen X.-C.,Clinical Medical Research Institute | And 12 more authors.
Clinical and Experimental Pharmacology and Physiology | Year: 2015

Adeno-associated virus (AAV) has become one of the most promising gene transfer tools for gene therapy. This work aims to evaluate tropism, gene transfer efficiency and safety of AAV9 vectors produced with recombinant baculovirus (rBac)-based system. AAV9-CMV-GFP and AAV9-CBA-GFP were produced using a rBac system, 1 × 1011 particles of each vectors were administered intravenously (i.v.) into mice and animals were killed at 1, 2, 3, 4, 5 and 8 weeks after administration. The GFP expression in different organs was analyzed by fluorescence imaging and Western blot. Viral genomic quantities were measured using qPCR. In vitro transduction efficiency of AAV9 vectors in primary cardiomyocytes and hepatocytes was determined by flow cytometry. Toxicity of AAV9 vectors was evaluated by determining certain cardiac and liver injury biomarkers and renal function test in vivo and TUNEL analysis in vitro. The data showed that AAV9 viral particles packaged by the rBac system were fully functional in vivo and in vitro. The CMV promoter predominantly induced higher cardiac GFP transgene expression and DNA copy numbers while the CBA promoter resulted in robust GFP expression and high vector DNA copy numbers in mouse liver, both in a time-dependent increased manner. Such distinct preferential effects were also observed in the heart and liver as early as 3 and 5 days after co-infection. Both the AAV9-CMV and AAV9-CBA viral packages did not induce heart, liver and renal damage and cell apoptosis. These results indicated that AAV9-CMV can efficiently and safely direct cardiac gene transfer, whereas AAV9-CBA is preferential for liver gene delivery. Copyright © 2015 Wiley Publishing Asia Pty Ltd.

Chen X.,Clinical Medical Research Institute | Chen B.,Clinical Medical Research Institute | Yang Y.,Clinical Medical Research Institute | Zhou Y.,Clinical Medical Research Institute | And 4 more authors.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology | Year: 2016

OBJECTIVE: To establish a simple, reliable and efficient isolation and culture method of human umbilical vein endothelial cells (HUVECs) in vitro.METHODS: Type 2 collagenase was used to digest umbilical cord and separate HUVECs. The cells were cultured in the endothelial cell culture medium (ECM). The cell morphology was observed under an inverted phase-contrast microscope. Immunofluorescence technique was applied to detect the expression of von Willebrand factor (vWF). Cell purity was determined by detecting CD31 level on cell surface with flow cytometry. Tube formation assay was used to test the function of the endothelial cells after cryopreservation in vitro.RESULTS: HUVECs successfully isolated were proved with high purity and good activity. HUVECs of primary generation could merge into a single layer one week after isolation. The cells showed a typical cobblestone-like arrangement. Immunofluorescence technique validated that the cells could widely express vWF and the expression frequency of CD31 was 93.1%. The cells were still capable of forming the lumen structure after cryopreservation, indicating that the standardized cryopreservation method could well maintain the cell function.CONCLUSION: This is a simple, reliable and efficient method of isolating and culturing HUVECs in vitro.

PubMed | Clinical Medical Research Institute
Type: Comparative Study | Journal: Annals of nutrition & metabolism | Year: 2014

This study aimed to identify the best predictor of metabolic syndrome (MetS) by comparing the predicting ability of various anthropometric and atherogenic parameters in the Chinese Han population in Xinjiang.A representative, cross-sectional sample of 5,757 Chinese Han adults were selected from the Cardiovascular Risk Survey conducted from October 2007 to March 2010. MetS prevalence, area under the curve (AUC), distance on the receiver operating characteristic curve and the cut-offs of each variable were compared for the presence of MetS.According to the criteria of the International Diabetes Federation (IDF), the Joint Interim Statement (JIS) and the Third Adult Treatment Panel (ATPIII), 32.1, 48.5, 39.3% of men and 38.0, 45.1, 44.9% of women had MetS in Xinjiang. According to the IDF criteria, the waist-to-height ratio (WHtR) had the highest AUC value in men (0.836) and women (0.837), with the optimal cut-off of 0.54 in men and 0.53 in women. According to both the JIS and ATPIII criteria, triglycerides/high-density lipoprotein cholesterol TG/HDL-C had the highest AUC value in men (0.830 and 0.833, respectively) and women (0.832 and 0.827, respectively), with the optimal cut-offs being 1.6 and 1.2 in men and 1.1 and 1.1 in women, respectively.WHtR was the best predictor of MetS according to the IDF criteria while TG/HDL-C was the best predictor of MetS according to the JIS and ATPIII criteria.

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