2.The Study of Silica Nanoparticles Modified as Gene Carriers Transfected Adult Human Epidermal Keratinocyte
Jianda ZHOU ; Chengqun LUO ; Yong CHENG
Journal of Chinese Physician 2001;0(10):-
Objective To study the ability of silica nanoparticles as gene carriers for adult human epidermal keratinocytes gene transfection.Methods The silica nanoparticles-DNA conjugated with the enhanced green fluorescence protein plasmid DNA(pEGFP-N_1) was transfected into adult human epidermal keratinocytes.The expression of green fluorescence protein was investigated in transfected keratinocytes by eletromicroscope examine and the efficiency of gene transfection was revealed.Results The silica nanoparticles-DNA complexes can be effectively transfected into adult human epidermal keratinocytes and the efficiency of gene transfection was about 20%~30%.Conclusion The silica nanoparticles can be used as DNA carriers for gene transfection,and can efficiency transfect the pEGFP-N_1 into adult human epidermal keratinocytes.
3.Changes of fibrinolytic status and coagulation function of peripheral blood at the acute stage in patients with intracerebral hemorrhage
Yong CHENG ; Xuejun DENG ; Guigui ZHOU
Journal of Clinical Neurology 1997;0(06):-
Objective To explore the changes of fibrinolytic status and coagulation function of peripheral blood at the acute stage in patients with intracerebral hemorrhage(ICH).Methods The platelet(PLT) count and mean platelet volume (MPV),the levels of plasma fibrinogen(Fib) and D-dimer(D-D) were detected at
4.Effect of α-melanocyte stimulating hormone on human dermal fibroblasts: preliminary identification of protein expression
Yong CHENG ; Dan YAN ; Zhiwei LIANG ; Haiyang ZHOU
Chinese Journal of Medical Aesthetics and Cosmetology 2011;17(3):208-211
Objective To evaluate the effect of this neuroendocrine hormone on protein expression by treating the human dermal fibroblasts with a-melanocyte stimulating hormone (α-MSH ).Methods Thehuman dermal fibroblasts was cultured, and the total protein of the fibroblasts were separated with immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). After Coomassie bright blue staining, gel images were acquired by Image-scanner and then analyzed with the PDQuest software. 2-DE maps of fibroblasts were established. Partial differently expressed protein spots were incised from gels and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and MSDB database searching by Mascot? software were used for protein identification. Results Well-resolved, reproducible 2-DE patterns of dermal fibroblasts treated with and without crMSH were obtained. 8 differently expressed protein spots were detected, among which 8 obtained peptide mass fingerprints (PMF) by MALDI-TOF-MS analysis. Among these proteins, of particular interest were five proteins annexin I, HSP27 and lamin A, etc. Conclusions Proteins expressed by human dermal fibroblasts treated with or without crMSH are different, and some of the differently expressed proteins involve apoptosis, intracellular signal transduction and framework construction and so on, which may be associated with anti-fibrosis effects of (a)-MSH on human dermal fibroblasts.
5.Effects of Aminoguanidine on Blood Brain Barrier and Brain Ischemic Damage during Reperfusion of Cerebral Ischemia in Rats
wen-an, WANG ; geng-fa, WANG ; yong-wei, ZHOU ; jie, CHENG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To evaluate the therapeutic effects of aminoguanidine(AG) on cerebral ischemia-reperfusion damage in rats. Methods The intravascular thread models with 2 h of occlusion and 22 h of reperfusion were made in the rats.The brain infarction size and the degree of blood brain barrier(BBB) disruption in the ischemic regions were evaluated by staining with 2,3,5-triphenyl tetrazolium chloride and observing with Evans blue fluorescence microscope.HE staining was utilized for observing neutrophil infiltration. Results The brain infarction(volume,) the area of BBB disruption and the degree of neutrophil infiltration were dramatically decreased in the treatment group as compared to the control group(P
6.Individual Identification of Cartilage by Direct Amplification in Mass Disasters
Chuanhai WANG ; Cheng XU ; Xiangqin LI ; Yong WU ; Zhou DU
Journal of Forensic Medicine 2017;33(3):281-283
Objective T o explore the effectiveness of direct am plification for the ST R analysis of carti-lage, and to accelerate the effectiveness of disaster victim identification. Methods E ighty-eight cartilage sam ples w ere directly am plified by Pow erPlex誖21 kit, and the results of genotyping w ere com pared w ith that obtained by the m agnetic beads m ethod. Results In 88 cartilage sam ples, the ST R genotypes w ere successfully detected from 84 sam ples by direct am plification and m agnetic beads m ethod, and both the results of genotyping by tw o m ethod w ere consistent. Conclusion D irect am plification w ith Pow er-Plex誖21 kit can be used for ST R genotyping of cartilages. T his m ethod is operated easily and prom ptly, w hich has a potential application in the individual identification of m ass disasters.
7.Targeted exogenous EGFP gene editing in caprine fetus fibroblasts by zinc-finger nucleases.
Yuguo YUAN ; Baoli YU ; Shaozheng SONG ; Feng ZHOU ; Liqing ZHANG ; Yingying GU ; Minghui YU ; Yong CHENG
Chinese Journal of Biotechnology 2013;29(11):1573-1580
Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals
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Base Sequence
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Cloning, Organism
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Electrophoresis
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Endonucleases
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genetics
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metabolism
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Fetus
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Fibroblasts
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metabolism
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Gene Knockout Techniques
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Gene Targeting
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methods
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Goats
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Green Fluorescent Proteins
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genetics
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Molecular Sequence Data
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Mutation
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Zinc Fingers
8.Preparation of the cDNA microarray on the differential expressed cDNA of senescence-accelerated mouse's hippocampus.
Xiao-Rui CHENG ; Wen-Xia ZHOU ; Yong-Xiang ZHANG
Chinese Journal of Biotechnology 2006;22(3):457-464
Alzheimer' s disease (AD) is the most common form of dementia in the elderly. AD is an invariably fatal neurodegenerative disorder with no effective treatment. Senescence-accelerated mouse prone 8 (SAMP8) is a model for studying age-related cognitive impairments and also is a good model to study brain aging and one of mouse model of AD. The technique of cDNA microarray can monitor the expression levels of thousands of genes simultaneously and can be used to study AD with the character of multi-mechanism, multi-targets and multi-pathway. In order to disclose the mechanism of AD and find the drug targets of AD, cDNA microarray containing 3136 cDNAs amplified from the suppression subtracted cDNA library of hippocampus of SAMP8 and SAMR1 was prepared with 16 blocks and 14 x 14 pins, the housekeeping gene beta-actin and G3PDH as inner conference. The background of this microarray was low and unanimous, and dots divided evenly. The conditions of hybridization and washing were optimized during the hybridization of probe and target molecule. After the data of hybridization analysis, the differential expressed cDNAs were sequenced and analyzed by the bioinformatics, and some of genes were quantified by the real time RT-PCR and the reliability of this cDNA microarray were validated. This cDNA microarray may be the good means to select the differential expressed genes and disclose the molecular mechanism of SAMP8's brain aging and AD.
Aging
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genetics
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metabolism
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Alzheimer Disease
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genetics
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metabolism
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Animals
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Disease Models, Animal
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Gene Expression
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Gene Expression Profiling
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Hippocampus
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metabolism
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Male
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Mice
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Models, Animal
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Oligonucleotide Array Sequence Analysis
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Reverse Transcriptase Polymerase Chain Reaction
9.Experimental study of selective portal vein embolization with the mixture of ZT glue and lipiodol
Shutong ZHUANG ; Bin CHEN ; Shubang CHENG ; Yong NI ; Rumin ZHOU ; Haibin GAO
Journal of Chinese Physician 2008;10(9):1196-1198
Objective To evaluate the feasibility of Selective Portal Vein Embolization(SPVE)in rabbits with the mixture of ZT glue and Lipiodol.Methods Sixteen white New Zealand rabbits were randomly divided into 2 groups:Group A,ZT glue:Lipiodol(1:2)mixture and Group B Lipiodol group.SPVE of left branch was performed in each group under digital subtraction angiography.The distribution feature of the embolic agents and the histopathology of liver in each group were observed.The weight ratio of the right lobe to the whole liver at the 30th day after SPVE were recorded and analyzed.Results Permanent embolization were occurred in group A.Recanalization was appeared in group B.Atrophy of the embolized lobes and compensatory hypertrophy of none-embolized lohes was,observed..The weight ratio of the right lobe to the whole liver Was 69.41±5.10% in group A.There was statistical difference between these two groups(P<0.05).Conclusion There were permanent embolization after SPVE with the mixture of ZT glue and lipiodol.SPVE induced atrophy of the embolized lobes of liver and compensatory hypertrophy of none-embolized lobes.
10.Preliminary study of endovascular treatment of anterior circulation multiple occlusions in acute ischemic stroke
Zhensheng LIU ; Cheng LI ; Wei WANG ; Longjiang ZHOU ; Yong SUN ; Xiongwei KUANG ; Xinjiang ZHANG
Chinese Journal of Radiology 2014;48(9):754-757
Objective To evaluate the safety and efficacy of the endovascular treatment of anterior circulation multiple occlusions (AMO) in acute ischemic stroke.Methods The clinical data of 10 patients with AMO treated by endovascular method from January 2011 to August 2013 were retrospectively analyzed.The proximal internal carotid artery (ICA) occlusion was treated using angioplasty in order to achieve ideal location of the guiding catheter.When necessary,stenting was performed after the reconstitution of the intracranial vessel.Recanalization was assessed according to the thrombolysis in cerebral ischemia (TICI) grade.Clinical prognosis was assessed using mRS at 3 months.The National Institutes of Health Stroke Scale (NIHSS) on admission and at discharge was compared using t test.Results The intracranial vessel was recanalized successfully (TICI ≥ 2b) in 9 cases and cervical carotid was stented in 8 cases.Adverse events were recorded in 3 patients,including one case of asymptomatic subarachnoid hemorrhage and two cases of symptomatic intra-cerebral hemorrhage.Mortality rate was 10 % (n=1).At the three-month follow up,mRS ≤ 2 was observed in five patients.The mean NIHSS scores was 15.7±2.2 on admission and 9.6±4.7at discharge,and the difference was statistic significant(t=2.86,P=0.02).Conclusion Endovascular therapy of AMO is technically feasible,and relatively safe and effective.