2.Measuring the expression of exogenous BDNF of NSCs after infection of supernatant from recombi-nant retrovirus pLXSN-BDNF
Xuemei ZHOU ; Huiping YUAN ; Donglai WU ; Ya WANG ; Zhuo WANG
Chinese Journal of Microbiology and Immunology 2009;29(9):786-791
Objective To develop neural stem cells(NSCs) which can stably express exogenous brain-derived neurotrophic factor(BDNF) in vitro. Methods NSCs from the subependymal zone of embry-onic day 14.5(E14.5) rat brain were purified by limiting dilution assay and then infected with supernatant of recombinant retrovirus pLXSN-BDNF and retrovirus pLXSN. The original copy numbers of exogenous gene templates from three groups NSCs(pLXSN-BDNF viral infection group, pLXSN viral infection group, control group) were detected by fluorescent quantitative PCR(FQ-PCR). ELISA assay was used for determining the protein contents of BDNF of supernatant from three groups NSCs for six days continually after seeded in 24-well plates in the same cell density. Results NSCs were purified successfully by limiting dilution assay.The original copy numbers of exogenous BDNF gene templates from pLXSN-BDNF viral infection group by FQ-PCR were (19.57±0.65) × 10~3 copies/μl, higher than those of another two groups(P < 0.05). The protein contents of BDNF of supernatant from NSCs of pLXSN-BDNF viral infection group was highest among three groups and compared with another two groups had statistical significance (P <0.05) . Conclusion The purified NSCs can be transduced exogenous BDNF successfully with supematant of recombinant retrovir-us pLXSN-BDNF which provide experimental evidences and laying foundations for further research of retinal transplantation and quantization investigation of gene therapy for optic nerve injury.
3.Effect of xuefu zhuyu decoction in inducing angiogenesis gene regulation of endothelial cell line ECV304.
Dong GAO ; Wen-yuan CHEN ; Li-ya WU
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(2):153-156
OBJECTIVETo study the acting mechanism of endothelial cell line ECV304 in regulating angiogenesis induced by Xuefu Zhuyu Decoction (XFZY).
METHODSThe angiogenesis effect of XFZY-contained serum (XFZY-CS) was confirmed by observing its impact on proliferation, cell cycle, migration of ECV304 and on vascular neogenesis in vitro. Then the effect of XFZY on various angiogenesis controlling factors was analyzed with gene chip microarray technique.
RESULTSTreatment of XFZY-CS in 2.5% concentration for 48 h showed evident actions of enhancing ECV304 activity, increasing cell numbers of S phase, inducing cell migration and promoting the in vitro angiogenesis. Meanwhile, expressions of four angiogenesis controlling genes were up-regulated and 10 were down-regulated.
CONCLUSIONThe angiogenesis mechanism of ECV304 induced by XFZY is complex, it shows a multi-pathway and multi-target feature.
Angiogenesis Inducing Agents ; Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; drug effects ; Female ; Gene Expression Regulation ; drug effects ; Male ; Neovascularization, Physiologic ; drug effects ; Rats ; Rats, Sprague-Dawley
4.Time-dependent change in expression of type-Ⅰ collagen protein gene during the development of restenosis after arterial injury
Ya-Wen ZHANG ; Guo-Yuan ZHANG ; Zong-Gui WU
Academic Journal of Second Military Medical University 2001;22(2):133-137
Objective: To observe the alteration of type- Ⅰ collagen protein gene expression after arterial injury and investigate its effect on the development of restenosis. Methods: Firstly, thee xperimental carotid arterial injury rabbit model was constructed. Then, Norther n blot, in situ hybridization and histomorphometric analysis were used to de tect the expression of procollagen mRNA and the accumulation of collagen protein 1,2,4 weeks after injury. Results: Type- Ⅰ collag en mRNA increased 1 week after injury, peaked 2 weeks later and decreased 4 week s later. The deposition of the collagen protein account for a high percentage o f space in neointima on histomorphometric analysis. Conclusion: Collagen protein may play an important role in the development of neointima and restenosis.
5.Germ-line epimutations and human cancer
Wu PU-YUAN ; Fan YI-MEI ; Wang YA-PING
Chinese Journal of Cancer 2009;28(12):1236-1242
Epimutations are errors in the normal process of epigenetic regulation which can result in aberrant transcriptional silencing of a normally active gene or reactivation of a normally silent gene.Epimutations are generally considered to be somatic events and to be confined in affected tissues.However, recent studies of patients with hereditary nonpolyposis colorectal cancer(HNPCC) have showed that allele-specific hypermethylation of CpG islands in the promoter region of the MLH1 gene, one of the causes of the tumor,existed in all the tissues examined.In addition,germ-line epimutations of other tumor suppressor genes(TSGs),such as MSH2 and BRCA1,have also been reported,demonstrating that epimutations might arise in the germline(during gametogenesis or early embryonic development).The role of germ-line epimutations might be as important as germ-line mutations in human disease.We reviewed the update on germ-line epimutations of TSGs including the possible mechanisms underlying germ-line epimutations,the possibility of transgenerational inheritance.and their impact on our understanding of human disease.
6.Morbidity of Malignant Hyperthermia in Central Core Disease Patients Undergoing Orthopedics
Shi-wen WU ; Wei-ya MA ; Sheng-yuan YU ; Dingguo SHEN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(5):427-428
ObjectiveTo investigate the incidence of skeletal and/or articular malformations in central core disease (CCD) patients, and their risk of malignant hyperthermia (MH) in orthopedics. Methods28 unrelated patients with central core disease collected in the past 23 years were analyzed retrospectively. Results22 patients showed limb muscle weakness, while 6 patients didn't show any clinical symptom. Skeletal and/or articular malformations appeared in 21 patients, including joint contracture in 11, joint dislocation in 9, scoliosis in 12, lordosis in 5 and thoracic deformity in 1. 6 of the 10 patients could not endure orthopedic operation because of MH attack or dubitable MH attack. Calcium-induced calcium release (CICR) test performed on other 8 patients with MH family history or MH attack showed all those patients were MH susceptibilities. ConclusionSkeletal and/or articular malformations are common in CCD, as well as MH attack in orthopedics.
7.Effects of Airborne Particles on Micronuclear Frequency of Human Binucleate Lymphocytes
Fu-Sheng YUAN ; Ya-Ping MA ; Wu-Hong ZHAO ; Al ET ;
Journal of Environment and Health 1992;0(04):-
Objective To study the effects of airborne particles exposure on the micronucleus frequency of human binucleate lymphocytes. Methods Airborne particles were collected at a residential area of Taiyuan city with classification air samplers.The organic substance was extracted by dichloromethane acetone and methanol in a Soxhlet apparatus. Four metals,Ni Pb Cd and Cr in airborne particles were extracted by 1∶1 nitric acid and determined by atomic absorption spectrophotometry.The mutagenicity of the extracts of airborne particles was studied with cytokinesis-locking assay. Results The content of airborne particles in the area was 0.791 9 mg/m3exceeded the related standard by 4.28 times. The small particles contained more metals elements and organic substances than the big particles. The extracts of airborne particles induced a significant increase in micronucleus production P
8.Two methods of hepatic vascular exclusion for hepatectomy
Wei-Ping ZHOU ; Ai-Jun LI ; Si-Yuan FU ; Ze-Ya PAN ; Yuan YANG ; Liang TANG ; Meng-Ehao WU
Chinese Journal of General Surgery 2000;0(12):-
Objective To compare the effects of hepatic vein occlusion with tourniquet and Satinsky clamp in reseeting liver tumor involving the second hepatic portal.Methods From Jan 2003 to Jun 2006,180 patients underwent major liver resection with the selective hepatic vascular exclusion (SHVE).According to methods of hepatic vein occlusion,they were divided into two groups:Occlusion with tourniquet(tourniquet group,n=95)and occlusion with Satinsky clamp(Satinsky clamp group,n= 85).In tourniquet group,the hepatic veins were encircled and occluded with tourniquet,and in Satinsky clamp group,the hepatic veins were not encircled and clamped directly with Satinsky clamp.Data regarding the intraoperative and postoperative courses of the patients were analyzed.Results There was no difference between the two groups regarding the operating time,ischemia time,intraoperative blood loss and postoperative complications rate.The dissecting time of hepatic veins was significantly shorter in Satinsky group(6.2?2.4 min vs 18.3?6.2 min).lu the tourniquet group,five hepatic veins(one fight hepatic vein and four common trunk of left-middle hepatic veins)could not be dissected and encircled because of the tumors involving the cava hepatic junction.Another patient's common trunk of left-middle hepatic vein was inadvertently lacerated during the dissection.Hepatic veins in these 6 patients were occluded with Satinsky clamp successfully.Conclusion Occlusion with Satinsky clamping is safer and easier procedure than tourniquets in the resection of liver tumor involving the second porta hepatis.
9.Isolation and identification of chemical constituents from Taxus cuspidta Sibe et Zucc.
Di WU ; Ya-Nan LI ; Li-Jun WU ; Hui-Yuan GAO
Acta Pharmaceutica Sinica 2010;45(11):1398-1401
Three compounds were isolated from the extract of Taxus cuspidta Sibe et Zucc with the column chromatography on silica gel and preparative HPLC methods. Their structures were identified according to the physicochemical properties and spectral analysis, and they were identified as (E)-1-methoxy-2-O-(p-coumaroyl)-myo-inositol (1), 2-deacetoxy-7beta, 9a, 10beta-trideacetyltaxinine J (2) and (3aS, 4aR, 6S, 8S, 8aS, 9R, 10R, 10aS)-benz[f]azulene-6, 8, 9, 10 (3H)-terol, 3a, 4, 4a, 5, 6, 7, 8, 8a, 9, 10-decahydro-10a-(1-hydroxyl-1-methylethyl)-1, 8a-dimethyl-5-methylene (3). Among them, compound 1 was a new compound, and compounds 2, 3 were two novel natural products.
Azulenes
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chemistry
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isolation & purification
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Coumarins
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chemistry
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isolation & purification
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Inositol
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analogs & derivatives
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chemistry
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isolation & purification
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Molecular Structure
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Plants, Medicinal
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chemistry
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Taxoids
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chemistry
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isolation & purification
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Taxus
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chemistry
10.Preliminary studies on the chemical characterization and antioxidant properties of acidic polysaccharides from Sargassum fusiforme.
Jing ZHOU ; Nan HU ; Ya-lin WU ; Yuan-jiang PAN ; Cui-rong SUN
Journal of Zhejiang University. Science. B 2008;9(9):721-727
In order to investigate the antioxidant properties of the polysaccharides from the brown alga Sargassum fusiforme, the crude polysaccharides from S. fusiforme (SFPS) were extracted in hot water, and the lipid peroxidation inhibition assay exhibited that SFPS possessed a potential antioxidant activity. Hence, two purely polymeric fractions, SFPS-1 and SFPS-2 were isolated by the column of DEAE (2-diethylaminoethanol)-Sepharose Fast Flow, with their molecular weights of 51.4 and 30.3 kDa determined by high performance gel permeation chromatography (HPGPC). They were preliminarily characterized using chemical analysis in combination of infrared (IR) and nuclear magnetic resonance (NMR) spectroscopies and found to contain large amounts of uronic acids and beta-glycosidical linkages. The antioxidant activities of these two SFPS fractions were evaluated using superoxide and hydroxyl radical-scavenging assays. The results show that the antioxidant ability of SFPS-2 was higher than that of SFPS-1, probably correlating with the molecular weight and uronic acid content.
Antioxidants
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chemistry
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Hydrogen-Ion Concentration
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Molecular Weight
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Pilot Projects
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Polysaccharides
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chemistry
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Sargassum
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metabolism