1.Advances of EC promoting vascularization in tissue-engineered bone
International Journal of Biomedical Engineering 2008;31(2):119-122
The vaseularization of tissue-engineered bone is the key problem which the development and employment of large sized tissue-engineered bone.The vascular endothelial cell has a great effect on promoting vascularization in tissue-engineered bone.Vascularizations fall into two modes of vaseulogenesis and angiogenesis according to differences in source of endothelial cells.Co-culture of osteoblasts and vascular endothelial cells has better result than single culture of each kind of cells.Different ways of improving the vascularization,such as searching for new source of vascular endothelial cell,co-culture and in vivo experiment are investigated to meet the challenge of bone tissue engineering.
2.Differentiation of rabbit bone marrow mesenchymal stem cells into vascular endothelial cells induced by in vitro combination of three kinds of cytokines
Chinese Journal of Tissue Engineering Research 2007;0(21):-
BACKGROUND: It has been verified vascular endothelial cells can propagate and differentiate into vessel by planted in the tissue-engineered material in vivo, but it may cause severe trauma when obtain vascular endothelial cells, and it has limited sources. OBJECTIVE: To investigate whether bone marrow mesenchymal stem cells (BMSCs) can differentiate into vascular endothelial cells by combined induction of vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF) and insulin-like growth factor (IGF)-1 by experimental observation. DESIGN, TIME AND SETTING: The control cell observation experiment was conducted at the Central Laboratory of First Affiliated Hospital, Liaoning Medical University from November 2006 to June 2007. MATERIALS: An 8-week-old Japanese big ear rabbit was used to isolate BMSCs. VEGF, bFGF and IGF-1 were purchased from Peprotech Company. METHODS: After anaesthesia, bone marrow was extracted from the rabbit. Enough BMSCs were harvested by adherence and trypsinization and randomized into two groups. BMSCs in the induction group were inoculated in DMEM supplemented with 10% fetal bovine serum (FBS), 10 ?g/L VEGF, 1 ?g/L bFGF and 2 ?g/L IGF-1 for 2 weeks. BMSCs in the control group were inoculated in DMEM containing 10% FBS. MAIN OUTCOME MEASURES: Morphological observation, nitric oxide (NO) content detected, immunohistochemistry staining of Ⅷ factor-related antigen and Weibel-Palade body under a electron microscope in BMSCs. RESULTS: Five days after induction, BMSCs were distributed in cluster, showing round. Non-induced BMSCs were evenly distributed, showing spindle or triangle. Fourteen days after induction, NO content in supernatant was significantly higher in the induction group than in the control group (t=3.75, P
3.Application of hypoxia inducible factor 1 in bone defect healing
Chinese Journal of Tissue Engineering Research 2007;0(50):-
Bone defect repair is a complicated process of connective tissue repair under the control of various cytokines.It is the ample blood supply which is a necessary condition to promote regeneration of fracture,especially bone defect and nonunion.Hypoxia inducible factor 1 can induce formation of neovascularization with perfect physiological function in the lesion of bone defect by regulating various gene expression,and thus provide nutritional support and favorable conditions to metabolism for different cells in the process of osteogenic differentiation and osteogenic activity to promote the healing of fracture.Both gene therapy and using hypoxia inducible factor 1 directly have an ability to promote neovascularization in the lesion of fracture.It is a hotspot at present that hypoxia inducible factor 1 in the field of bone tissue engineering is used to treat bone defect.Hypoxia inducible factor 1 transgenic therapy needs further research in the repairing process of bone defect.
4.Investigation Effect of Naofuqing Capsules on Senile Dementia on 30 Cases
Danping GAO ; Guihai CHEN ; Jun YANG ;
Chinese Traditional Patent Medicine 1992;0(04):-
Objective: To investigate the clinical effect of Naofuqing Capsules on senile dementia. Methods: 30 patients with senile dementia were treated with Naofuqing Capsules and the curative results were compared with that of the control group 30 cases. Results: The results showed that the integral titres of MMSE and HDS R of groups increased obviously after treatment( P 0.05). The total effective rates were 83.3% and 80.0%,respectively. No obvious difference was observed in clinical effect between two groups. Conclusion: Naofuqing Capsules possesses a definite therapeutic effect for senile dementia.
5.Treatment of anterior canal benign paroxysmal positional vertigo by Yacovino repositioning maneuver.
Danping CHEN ; Shanshan XIONG ; Yong CUI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(1):17-19
OBJECTIVE:
To evaluate the efficacy of Yacovino repositioning maneuver in patients with anterior semicircular canal benign paroxysmal positional vertigo (ASC-BPPV).
METHOD:
Nine patients were diagnosed as ASC-BPPV from January 2013 to October 2014. All the patients were performed with Yacovino repositioning maneuver and the effective rate were evaluated by Dix-Hallpike tests.
RESULT:
Among the nine ASC-BPPV patients, 2 cases were successfully controlled by the first maneuver, 2 cases by the second time, and the nystagmus of 1 case was disappeared after 1 months' follow-up. The remaining 3 cases were respectively followed up till 7,8, 12 months with consistent positional downbeat nystagmus.
CONCLUSION
Being a relative low incidence disease, of ASC-BPPV also has low effective rate after Yacovino repositioning maneuver.
Benign Paroxysmal Positional Vertigo
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therapy
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Humans
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Patient Positioning
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Semicircular Canals
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Vertigo
6.Systematic Assessment on Effects of Wendan Decoction on Dyslipidemia
Xuanjing CHEN ; Xiaoguang CHEN ; Danping XU ; Huanlin WU
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(8):93-98
Objective To systematically assess the clinical efficacy and security of Wendan Decoction in the treatment of dyslipidemia. Methods Retrieved from CNKI, CBM, VIP database, Wanfang Database, and PubMed, articles of RCTs about Wendan Decoction in the treatment of dyslipidemia were included. The searching range was from the establishment of the database to April, 2016. NoteExpress3.2.0 was used to establish database. Two researchers independently screened the articles and extracted the data. Cochrane Systematic Review Manual 5.1.0 was used for quality evaluation. Meta-analysis was performed with RevMan5.3 software. Results 12 articles involving 848 patients were included. Results of Meta-analysis showed that compared with the western medicine treatment, Wendan Decoction group was more capable of significantly improving clinical effective rate of dyslipidemia [OR=2.46, 95%CI (1.41, 4.30), P=0.001], lowering the level of TG [WMD=-0.31, 95%CI (-0.46, -0.15), P=0.0001] and LDL-C [WMD=-0.25, 95%CI (-0.30, -0.20), P=0.00001], but there was no statistical significance in regulating TC [WMD=-0.14, 95%CI (-0.35, -0.08), P=0.49] and HDL-C [WMD=0.04, 95%CI (-0.01, -0.1), P=0.13] between the two groups; compared with the western medicine group, Wendan Decoction combined with western medicine group was more capable in the clinical effective rate of dyslipidemia (P<0.05), regulating the level of TG [WMD=0.30, 95%CI (-0.37, -0.23), P=0.00001], TC [WMD=0.65, 95%CI (-0.80, -0.69), P=0.00001], LDL-C (P<0.05, P<0.01) between the two groups; Wendan Decoction combined with western medicine group had low rate of adverse reactions. Conclusion Wedan Decoction can effectively regulate dyslipidemia, with good safety. High quality RCTs in large-scale and multi-center are necessary for further evidence due to the poor methodological quality of included trials with few samples and other limitations.
8.Construction of adenovirus vectors carrying VEGF121-FLAG and hrGFP-1 and their expressions in bone marrow stromal stem cells
Danping LIU ; Chen LI ; Liang HU ; Guoxian WANG
Chinese Journal of Tissue Engineering Research 2010;14(45):8539-8543
BACKGROUND: Vascular endothelial growth factor (VEGF) can promote angiogenesis, and has been extensively used in treatment of bone defect. However, few studies have addressed its isomer VEGF121. OBJECTIVE: To construct adenovirus vector carrying VEGF121-FLAG and humanized Renilla reniformis green fluorescent protein 1(hrGFP-1) and observe its expression in bone marrow stromal stem cells (BMSCs). METHODS: Using polymerase chain reaction technique, VEGF121 gene contained in the plasmid of pTG19T-VEGF121 was used to remove termination codon. NotI and Xho I restriction sites were added before and after gene sequence. Obtained gene subclone was moved onto pMD19-T plasmid. The pMD19-T-VEGF121 and pShuttle-CMV-IRES-hrGFP-1 plasmids underwent double enzymatic digestion. Small fragment and big fragment were retrieved utilizing gel. Subsequently, coupled reaction was conducted to complete the construction of shuttle plasmid. After measuring virus titer, BMSCs were transfected and the fluorescence intensity was observed under fluorescence microscope.RESULTS AND CONCLUSION: Recombinant adenovirus plasmid was successfully constructed by enzymatic digestion determination and gene sequence. Fluorescence microscope has shown that BMSCs transfected with recombinant adenovirus presented significantly green fluorescence expression. Thus, adenovirus vector carrying VEGF121-FLAG and hrGFP-1 gene can express in eukaryotic cells, which can be used for gene therapy for ischemic disease.
9.Construction of adenovirus-mediated eukaryotic expression vector co-expressing mutant hypoxia-inducible factor-1 alpha target protein and humanized Renilla reniformis green fluorescent protein reporter molecule under normoxic conditions
Danping LIU ; Guoxian WANG ; Liang HU ; Chen LI
Chinese Journal of Tissue Engineering Research 2010;14(20):3787-3792
BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) can regulate the co-expression of various genes, and can induce angiogenesis with integrated physiological function.OBJECTIVE: To construct a novel adenoviral eukaryotic expression vector that can co-express mutant hypoxia-inducible factor-1 alpha (HIF-1a) target protein and humanized Renilla reniformis green fluorescent protein (hrGFP) reporter molecule under normoxic conditions.METHODS: The human HIF-1α gene carried by target gene donor plasmid pCMV6-XL5-HIF1α was sequenced and the site of restriction enzyme in above gene was analyzed. Site-directed mutagenesis of three amino acids including the 402 location, the 564 location, and the 803 location in gene coding region in HIF-1α were performed by polymerase chain reaction and sequencing was also done for monitoring mutation. The HIF-1α gene mutated correctly (HIF-1αmu) was coupled to adenoviral shuttle vector pShuttle-CMV-IRES- hrGFP-1. The recombinant adenovirus shuttle vector carrying HIF-1αmu gene was transferred to BJ5183-AD-1 electroporation competent cells after sequencing identification and Pme I restriction enzyme linearization.HIF-1αmu and hrGFP gene as well as hemeo-expression elements of hrGFP gene were reconstructed into adenoviral genome plasmids using homologous recombination mechanism in bacterium. Recombinants were obtained by Pac I restriction enzyme digestion and sequencing identification.RESULTS AND CONCLUSION: Amino acids including the 402 location, the 564 location and the 803 location in gene coding region in HIF-1α had become alanine after site-directed mutagenesis. Recombinant adenoviral expressing vector was successful as confirmed by restriction enzyme digestion and sequencing. These findings demonstrate that a novel recombinant adenoviral mutant eukaryotic expression vector pAd-HIF1αmu-IRES-hrGFP-1 was successfully constructed.
10.One case of late-onset adrenal hypoplasia congenita caused by a novel mutation of DAX-1 gene
Danping WANG ; Cunren CHEN ; Yanxia LIU ; Lijuan WANG ; Xialian LI
Chinese Journal of Endocrinology and Metabolism 2011;27(1):47-49
A novel hemizygous frameshift mutation in exon1of DAX-1 gene (993delC) was found in a patient with late-onset adrenal hypoplasia congenita and hypogonadotropic hypogonadism.This mutation led the stop codon to appear in advance of 59 amino acids.His mother and two sisters were the carriers of this hemizygous mutation while his father and brother were wild-type.After glucocorticoid hormone replacement therapy, the clinical symptom was improved, but the level of ACTH was not suppressed.