5.Biocompatibility of bone marrow mesenchymal stem cells with bladder acellular matrix scaffold
Xiaojun ZHAO ; Jun YU ; Yingfei DUAN
Chinese Journal of Tissue Engineering Research 2015;(36):5769-5773
BACKGROUND:In the repair of urinary tract defects, we have been actively trying to construct the urinary tract substitutes with normal physiological function through combining ideal seed cel s and proper scaffold materials by tissue engineering method. OBJECTIVE:To investigate the biocompatibility of bone marrow mesenchymal stem cel s with rabbit bladder acel ular matrix scaffold. METHODS:Rabbit bone marrow mesenchymal stem cel s were isolated and cultured using density gradient centrifugation method. Passage 3 rabbit bone marrow mesenchymal stem cel s were cultured on the rabbit bladder acel ular matrix. The cel s were counted every day for 12 days, to drawn a cel growth curve. Bone marrow mesenchymal stem cel s cultured alone were used as control group. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cel s were successful y seeded onto the bladder acel ular matrix. Under the inverted microscope, the cel s grew out of the bladder acel ular matrix, and a great amount of long spindle-shaped cel s were found around the bladder acel ular matrix. With 5 days of inoculation, the cel s in the two groups grew gently;at 6-9 days, the cel growth curve gradual y became steeper, and the cel division and growth were increased exponential y;at 10-12 days, the cel s recovered to a gentle state. Cel growth curves in the two groups were basical y coincident, suggesting that rabbit bone marrow mesenchymal stem cel s have good biocompatibility with the bladder matrix.
6.The myocardial injury after limb ischemia/reperfusion in rats and the protective effects of NO on myocardium.
Li-Jun ZHAO ; Guo-Xian DUAN ; Shu-Yun DONG
Chinese Journal of Applied Physiology 2007;23(3):268-327
Animals
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Extremities
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blood supply
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Ischemia
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metabolism
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Male
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Myocardial Reperfusion Injury
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metabolism
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Nitric Oxide
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blood
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Oxidative Stress
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Rats
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Rats, Wistar
7.Effects of SREBP-1 targeted RNAi on lipid droplet formation in HKC cells under stimulation of high glucose
Jun HAO ; Hong SHI ; Song ZHAO ; Yunzhuo REN ; Huijun DUAN
Chinese Pharmacological Bulletin 1987;0(03):-
Aim To construct eukaryotic expression vector of shRNA(small hairpin RNA)for human SREBP-1(sterol regulation element binding protein-1)gene and explore its effects on lipid droplet formation in human renal proximal tubular epithelial cell line(HKC)under the stimulation of high glucose.Methods Two eukaryotic expression vectors of shRNA were constructed for human SREBP-1 gene.The HKC cells were transfected with negative control plasmid(pGenesil-1-HK)and two recombinant vectors(pGenesil-1-SREBP1-1 and pGenesil-1-SREBP1-2)and then were cultured under the stimulation of high glucose for about 48 h.The expression of SREBP-1 mRNA and FAS mRNA was detected by RT-PCR and SREBP-1 protein expression was investigated by Western blot.Lipid droplets were detected by Oil Red O staining.Results DNA sequencing showed that the target segments were successfully cloned into pGenesil-1 vector respectively.RT-PCR indicated that two recombinant vectors could inhibit the expression of SREBP-1 mRNA and FAS mRNA in HKC cells under the stimulation of high glucose.Similarly,SREBP-1 protein was also inhibited by the transfection with recombinant vectors.Oil Red O staining found that silencing of SREBP-1 gene resulted in lipid droplets decrease.Conclusions The eukaryotic expression vector of shRNA for human SREBP-1 gene was successfully constructed,and the expression of SREBP-1 was inhibited effectively by the expressed siRNA in HKC cells that resulted in lipid droplets decrease through FAS mRNA transcription inhibition.
8.The significance of NO in renal cell apotosis following hind limbs ischemia/reperfusion in rats.
Guo-xian DUAN ; Lian-yuan ZHAG ; Li-jun ZHAO
Chinese Journal of Applied Physiology 2009;25(2):281-285
Animals
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Apoptosis
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Extremities
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blood supply
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Kidney
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pathology
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Male
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Nitric Oxide
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physiology
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Random Allocation
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Rats
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Rats, Wistar
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Reperfusion Injury
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pathology
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physiopathology
9.A review of detection methods for human bocaviruses.
Yan LU ; Dan-Di LI ; Yu JIN ; Zhao-Jun DUAN
Chinese Journal of Virology 2014;30(3):298-302
Human bocavirus (HBoV) 1-4 have been detected both in respiratory and stool samples since the first HBoV was discovered in 2005. HBoV-1 is mostly associated with respiratory infection, while HBoV 2-4 are usually associated with intestinal tract infection. A variety of signs and symptoms have been described in patients with HBoV infection, including cough, wheezing, pneumonia, and diarrhea, but the research on pathogenic mechanism of HBoV is limited because HBoV cannot be cultured in vitro due to the lack of appropriate host cells. Three-dimensional epithelial cell culture, reverse genetics, and viral metagenomics are identified as novel tools that may promote the research on pathogenic mechanism of HBoV and the discovery of new viruses. This review summaries currently available diagnostic approaches such as electron microscopy, cell culture, PCR, and immunoassay in order to provide a method reference for indepth research on HBoV.
Animals
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Human bocavirus
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genetics
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growth & development
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isolation & purification
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pathogenicity
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Humans
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Parvoviridae Infections
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diagnosis
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virology
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Viral Proteins
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genetics
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metabolism
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Virology
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methods
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Virulence
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Virus Cultivation
10.Research progress in human adenovirus.
Wen-Juan GAO ; Yu JIN ; Zhao-Jun DUAN
Chinese Journal of Virology 2014;30(2):193-200
Human adenovirus (HAdV) is one of the most important pathogens in infants and young children with acute respiratory infections and other diseases. This article reviews the literature on HAdV, including its molecular biological characteristics, detection and typing, and pathogenic mechanism, the clinical features and epidemiological characteristics of HAdV-related diseases, and the prevention and control of HAdV infections. So far, 67 types of HAdV have been identified, including recombinant variants discovered in recent years. The major epidemic strains that cause acute respiratory infections are HAdV-3 and HAdV-7, both of which belong to the subgroup B. HAdV often leads to acute respiratory infections, but it also causes diseases of other systems. HAdV-related diseases have similar clinical manifestations as those caused by other respiratory viruses, but often accompanied by gastrointestinal symptoms. The pathogenic mechanism of HAdV remains unclear, especially for the new recombinant variants, due to few studies on their association with diseases. Because there are no prospective, large randomized controlled trials of HAdV infections, the treatment of HAdV infections is controversial. Vaccine is the most effective measure to reduce respiratory HAdV infections, but it is still not commercially available.
Adenovirus Infections, Human
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virology
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Adenoviruses, Human
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classification
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genetics
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isolation & purification
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physiology
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Animals
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Humans