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Teachers as Collaborators to Motivate Learning Autonomy in English Speaking Class--Two Cases at a Senior High School and a University
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作者 Yiling Chen 《通讯和计算机(中英文版)》 2012年第8期905-908,共4页
关键词 自主学习能力 英语口语 作者 教师 学生 大学 中和 激励
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Ⅱ型糖尿病兔大脑皮质和海马神经元的神经丝蛋白表达(英文) 被引量:1
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作者 马志健 刘正清 +4 位作者 张秋菊 蔡维君 李明波 刘小丹 陈二云 《中国医学工程》 2002年第6期17-20,共4页
Objective:To investigate the morphological changes of the neuronal neurites in diabetic rabbit brain. Methods: Twenty- four New Zealand White rabbits were divided into 2 groups: control group and type Ⅱ diabetic grou... Objective:To investigate the morphological changes of the neuronal neurites in diabetic rabbit brain. Methods: Twenty- four New Zealand White rabbits were divided into 2 groups: control group and type Ⅱ diabetic group induced by high - carbohydrate and high- fat diet. The levels of blood sugar and insulin were detected at week 0(w0), w4, w8, w13, w18, w23 and w28. Brain tissue was stained by Nissl staining and immunolistochemistry with a specific antibody to neurofilament proteins. Result: In diabetic rabbits, the amount of large pyramidal neuron was significantly reduced, and neuronal neurites became swollen, whorled, disrupted and changed in caliber. In hippocampus CA1 region neurofilament staining was very weak. Conclusion: Neurotoxicity of chronic hyperglycemia might be relevant to vascular chronic complications, which affected the expression of NF and led to neurophysiological and structural changes in the brain of rabbits with type Ⅱ diabetes. 展开更多
关键词 New Zealand White Rabbits Type Diabetes NEUROFILAMENT Diabetic Encephalopathy
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Interactions between mycoplasma lipid-associated membrane proteins and the host cells 被引量:18
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作者 YOU Xiao-xing ZENG Yan-hua WU Yi-mou 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2006年第5期342-350,共9页
Mycoplamas are a group of wall-less prokaryotes widely distributed in nature, some of which are pathogenic for humans and animals. There are many lipoproteins anchored on the outer face of the plasma membrane, called ... Mycoplamas are a group of wall-less prokaryotes widely distributed in nature, some of which are pathogenic for humans and animals. There are many lipoproteins anchored on the outer face of the plasma membrane, called lipid-associated membrane proteins (LAMPs). LAMPs are highly antigenic and could undergo phase and size variation, and are recognized by the innate immune system through Toll-like receptors (TLR) 2 and 6. LAMPs can modulate the immune system, and could induce immune cells apoptosis or death. In addition, they may associate with malignant transformation of host cells and are also con-sidered to be cofactors in the progression of AIDS. 展开更多
关键词 MYCOPLASMA Lipid-associated membrane proteins (LAMPs) Toll-like receptor (TLR) Immunomodulin
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Synthesis and Anticancer Effect of gem-Difluoromethylenated Chrysin Derivatives 被引量:21
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作者 Xing ZHENG Jian Guo CAO +2 位作者 Duan Fang LIAO Bing Yang ZHU Hui Ting LIU 《Chinese Chemical Letters》 SCIE CAS CSCD 2006年第11期1439-1442,共4页
Ten gem-difluoromethylenated chrysin derivatives were prepared and their anticancer activities in vitro were evaluated by the standard MTT method. The results of biological test showed that some of gem-difluoromethyle... Ten gem-difluoromethylenated chrysin derivatives were prepared and their anticancer activities in vitro were evaluated by the standard MTT method. The results of biological test showed that some of gem-difluoromethylenated chrysin derivatives had higher anticancer activity than chrysin. 展开更多
关键词 Anticancer activity gem-difluoromethylenated chrysin.
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Role of NLRP3 and NLRP1 inflammasomes signaling pathways in pathogenesis of rheumatoid arthritis 被引量:12
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作者 Tao Wang Chang-Liang Zhu +4 位作者 Shuai Wang Li-Wen Mo Guo-Dong Yang Jun Hu Fan Zhang 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2014年第10期827-831,共5页
Objective:To investigate the role of NLRP3 and NLRP1 inflammasomes signaling pathways in rheumatoid arthritis(RA).Methods:A total of 36 patients with RA were selected,peripheral blood mononuclear cell(PBMC)and granulo... Objective:To investigate the role of NLRP3 and NLRP1 inflammasomes signaling pathways in rheumatoid arthritis(RA).Methods:A total of 36 patients with RA were selected,peripheral blood mononuclear cell(PBMC)and granulocyte were separated from venous blood.RT-qPCR method was used to detect the expression level and diversity of NLRP3 and NLRP1 in PBMC and granulocyte mRNA in patieuts with RA.and detect the mRNA expression of downstream factor IL-1β.The correlation between RA and the expression of NLRP3 aud NLRP1 was analyzed.Normal 30 cases were set as control group.Results:Expression levels of NLRP1.and caspase-1mRNA in PBMC of RA group were significautly lower than those of control group(P<0.05).while there was no significant differeuee in expression levels of NLRP3,ASC.IL-1βmRNA between these two groups(P>0.05);NLRP3,caspase-1,and ASC mRNA expression in granulocyte of RA patients were significantly lower than those in control group(P<0.05).There was no currelation between rheumatoid factor and expression levels of NLRP3.ASC.caspase-1 mRNA in RA group(P>0.05);NLRP1,IL-1βmRNA expression level had a negative corrlation with anti-rheunatoid factor antibody(P=0.0332,0.0340).Conclusions:NLRP3 and NLRP1 inflammasomes signaling pathways are involved in RA inflammatory reaction process as protective factors,and play an important role in RA inflammatory mechanisms. 展开更多
关键词 NLRP3 NLRP1 RHEUMATOID ARTHRITIS Immune mechanism
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Opti mization and Model of Laminar Cooling Control Systemfor Hot Strip Mills 被引量:22
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作者 XIE Hai-bo LIU Xiang-hua +1 位作者 WANG Guo-dong ZHANG Zhong-ping 《Journal of Iron and Steel Research International》 SCIE EI CAS CSCD 2006年第1期18-22,共5页
The structure of laminar cooling control system for hot rolling was introduced and the control mode, cooling strategy, segment tracking and model recalculation were analyzed. The parameters of air/water cooling models... The structure of laminar cooling control system for hot rolling was introduced and the control mode, cooling strategy, segment tracking and model recalculation were analyzed. The parameters of air/water cooling models were optimized by regressing the data gathering in situ, and satisfactory effect was obtained. The coiling temperature can be controlled within ±15℃. 展开更多
关键词 hot rolled strip laminar cooling control system model parameter optimization
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Effects of extract of Buddleja officinalis on partial inflammation of lacrimal gland in castrated rabbits with dry eye 被引量:15
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作者 Xiao-Lei Yao, Hui-An Zhu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2010年第2期114-119,共6页
AIMTo assess the effects of extract of Buddleja officinalis on tear secretion volume, tear film stability, expressions of TGF-β1, IL-1β, TNF-α in lacrimal gland of castrated rabbits with dry eye.
关键词 dry eye lacrimal gland extract of buddleja officinalis ANDROGEN
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Evaluation of novel decellularizing corneal stroma for cornea tissue engineering applications 被引量:15
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作者 Yi Shao, Lu Yang 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2012年第4期415-418,共4页
AIM:To develop a new decellularization method depended upon the natural corneal structure and to harvest an ideal scaffold with good biocompatibilities for corneal reconstruction.METHODS:The acellular cornea matrix (A... AIM:To develop a new decellularization method depended upon the natural corneal structure and to harvest an ideal scaffold with good biocompatibilities for corneal reconstruction.METHODS:The acellular cornea matrix (ACM) were prepared from de-epithelium fresh porcine corneas (DFPCs) by incubation with 100% fresh human sera and additional electrophoresis at 4℃. Human corneal epithelial cells (HCEs) were used for the cytotoxicity tests of ACM. ACM were implanted into the Enhanced Green Fluorecence Protein (eGFP) transgenic mouse anterior chamber for evaluation of histocompatibility.RESULTS:HE and GSIB4 results showed fresh porcine cornea matrix with 100% human sera and electrophoresis could entirely decellularize stromal cell without reducing its transparency. ACM has no cytotoxic effect ex vivo. Animal test showed there was no rejection for one month after surgery.CONCLUSION:These results provide a decellularizing approach for the study of corneal tissue engineering and had the broader implications for the field of biological tissue engineering in other engineered organ or tissue matrix. 展开更多
关键词 CORNEA human sera ELECTROPHORESIS tissue engineering DECELLULARIZATION BIOCOMPATIBILITY
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Investigation of transcriptional gene silencing and mechanism induced by shRNAs targeted to RUNX3 in vitro 被引量:2
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作者 Xue-Zhi Feng Xiu-Sheng He +6 位作者 Ying-Zhi Zhuang Qiao Luo Jun-Hao .liang Shuai Yang Xue-Fang Tang Ju-Lin Liu Tao Chen 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第19期3006-3014,共9页
AIM:To investigate transcriptional gene silencing induced by short hairpin RNAs (shRNAs) that target gene prompter regions of RUNX3 gene, and whether shRNAs homologous to DNA sequences may serve as initiators for meth... AIM:To investigate transcriptional gene silencing induced by short hairpin RNAs (shRNAs) that target gene prompter regions of RUNX3 gene, and whether shRNAs homologous to DNA sequences may serve as initiators for methylation. METHODS:According to the principle of RNAi design, pSilencer3.1-H1-shRNA/RUNX3 expression vector was constructed, The recombinant plasmid shRNA was transfected into human stomach carcinoma cell line SGC7901 with Lipofectamine 2000. Then, the positive cell clones were screened by G418. The mRNA and protein expression level of RUNX3 in the stable transfected cell line SGC7901 were determined by RT-PCR, Western blotting and immunocytochemistry. Characteristics of the cell lines including SGC7901, pSilencer3.1-H1/SGC7901 and pSilencer3.1-H1-shRNA/RUNX3/SGC7901 were analyzed with growth curves, clone formation rate and cell-cycle distribution. The activated level of RUNX3 was examined after treatment with the different density of 5’-aza-2’-deoxycytidine (5-Aza-CdR) by using semi- quantitative RT-PCR and Western blotting. RESULTS:In the cell line SGC7901 transfected with pSilencer3.1-H1-shRNA/RUNX3, mRNA and proteinexpression of the RUNX3 gene was lost identified by RT- PCR, Western blotting and immunocytochemistry assay. The growth of pSilencer3.1-H1-shRNA/ RUNX3/SGC7901 cells without expression of RUNX3 was the fastest (P < 0.05), its rate of clone formation was the highest (P < 0.01), and the cell distribution in G0/G1 and S/M phases was lowest and highest, respectively (P < 0.05), compared with that of the transfected pSilencer3.1-H1 and non-transfected cells. Through RT-PCR and Western blot assay, inactivated RUNX3 could not be reactivated by 5-Aza-CdR. CONCLUSION:We found that, although shRNAs targeted to gene prompter regions of RUNX3 could effectively induce transcriptional repression with chromatic changes characteristic of inaction promoters, this was independent of DNA methylation, and the presence of RNA-dependent transcriptional silencing showed that RNA-directed DNA methylation might be an existing gene regulatory mechanism relative to the methylated in humans. 展开更多
关键词 RNA interference Short hairpin RNAs PROMOTER DNA methylation RUNX3 Stomach carcinoma
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Artificial neural network based inverse design method for circular sliding slopes 被引量:4
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作者 丁德馨 张志军 《Journal of Central South University of Technology》 EI 2004年第1期89-92,共4页
Current design method for circular sliding slopes is not so reasonable that it often results in slope (sliding.) As a result, artificial neural network (ANN) is used to establish an artificial neural network based inv... Current design method for circular sliding slopes is not so reasonable that it often results in slope (sliding.) As a result, artificial neural network (ANN) is used to establish an artificial neural network based inverse design method for circular sliding slopes. A sample set containing 21 successful circular sliding slopes excavated in the past is used to train the network. A test sample of 3 successful circular sliding slopes excavated in the past is used to test the trained network. The test results show that the ANN based inverse design method is valid and can be applied to the design of circular sliding slopes. 展开更多
关键词 circular sliding slopes artificial neural network inverse design
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BINDING AND INTERACTION OF TRANSCRIPTIONAL REGULATOR HUMAN DAXX WITH ADENOVIRUS12 E1B 55 KILODALTON ONCOPROTEIN 被引量:2
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作者 万艳平 吴移谋 +3 位作者 朱翠明 谭立志 余敏君 廖端芳 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2003年第4期273-276,共4页
Objective: To study the disruption of co- localization of human Daxx(hDaxx) with promyelocytic leukemia protein(PML) at the PML oncogenic domains (PODs) by the interaction of hDaxx with adenovirus(Ad) 12 E1B 55 Kiloda... Objective: To study the disruption of co- localization of human Daxx(hDaxx) with promyelocytic leukemia protein(PML) at the PML oncogenic domains (PODs) by the interaction of hDaxx with adenovirus(Ad) 12 E1B 55 Kilodalton Oncoprotein (Ad12 E1B 55kD). Methods: The direct binding reaction of hDaxx and Ad12 E1B 55kD was analyzed by coimmunoprecipitation and Western blotting in vivo or in vitro. The interaction of hDaxx with Ad12 E1B 55kD was studied using yeast two-hybrid assay. Results: hDaxx bounded directly to Ad12 E1B 55kD in vivo and in vitro. hDaxx interacted with full length Ad12 E1B 55kD. Conclusion: Transcriptional regulator hDaxx directly binds to and interacts with Ad12 E1B 55kD. 展开更多
关键词 Human Daxx Ad12 E1B 55kD BINDING INTERACTION
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Resonance Light Scattering Spectra of Silver Thiocyanate System and its Application in Analysis 被引量:2
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作者 Chang Yin LU Zhi Hui HAN Sheng Yuan YANG 《Chinese Chemical Letters》 SCIE CAS CSCD 2005年第8期1063-1066,共4页
A novel determination method of Ag^+ was established. In acetic acid-sodium acetate buffer (pH 5.0) medium, Ag^+ reacts with SCN^- to form AgSCN in the presence of TritonX-100,which results in an increase of reson... A novel determination method of Ag^+ was established. In acetic acid-sodium acetate buffer (pH 5.0) medium, Ag^+ reacts with SCN^- to form AgSCN in the presence of TritonX-100,which results in an increase of resonance light scattering (RLS)and giving a new RLS spectrum.The maximum RLS peak was at 585 nm,The enhancement of resonance light scattering at 585 nm was proportional to the concentration of Ag^+ ranging from 0.0045-4.00μg mL^-1 (r=9991),and the detection limit was 1.37 ng mL^-1 with the recovery of 97.70%- 104.80%。 展开更多
关键词 Resonance light scattering SILVER silver thiocyanate.
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Benign symmetric lipomatosis (Madelung’s disease) with concomitant incarcerated femoral hernia: A case report 被引量:2
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作者 Bo Li Zheng-Xing Rang +2 位作者 Jia-Cong Weng Guo-Zuo Xiong Xian-Peng Dai 《World Journal of Clinical Cases》 SCIE 2020年第21期5474-5479,共6页
BACKGROUND Benign symmetric lipomatosis(BSL)was first described by Brodie in 1846 and defined as Madelung’s disease by Madelung in 1888.At present,about 400 cases have been reported worldwide.Across these cases,surgi... BACKGROUND Benign symmetric lipomatosis(BSL)was first described by Brodie in 1846 and defined as Madelung’s disease by Madelung in 1888.At present,about 400 cases have been reported worldwide.Across these cases,surgical resection remains the recommended treatment.Here we report a case of neck BSL with concomitant thick fatty deposit in the inguinal region,which concealed the signs of a right incarcerated femoral hernia.CASE SUMMARY A 69-year-old male patient was admitted to our hospital with“abdominal pain,abdominal distension,nausea-vomiting and difficult defecation for half a month”.Moreover,he had a mass in the right inguinal region for more than 10 years.An egg-sized neck mass also developed 15 years ago and had developed into a full neck enlargement 1 year later.In addition,the patient had a history of heavy alcohol consumption for more than 40 years.With the aid of computerized tomography scan,the patient was diagnosed with BSL and a low intestinal mechanical obstruction caused by a right inguinal incarcerated hernia.Under general anesthesia,right inguinal incarcerated femoral hernia loosening and tension-free hernia repair was performed.However,this patient did not receive BSL resection.After a 1-year follow-up,no recurrence of the right inguinal femoral hernia was found.Moreover,no increase in fat accumulation was found in the neck or other areas.CONCLUSION Secretive intraperitoneal fat increase may be difficult to detect,but a conservative treatment strategy can be adopted as long as it does not significantly affect the quality-of-life. 展开更多
关键词 Benign symmetric lipomatosis Madelung’s disease Neck Inguinal region Inguinal incarcerated hernia Case report
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Oscillation of Nonlinear Impulsive Delay Hyperbolic Partial Differential Equations 被引量:2
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作者 罗李平 彭白玉 欧阳自根 《Chinese Quarterly Journal of Mathematics》 CSCD 2009年第3期439-444,共6页
In this paper,by making use of the calculous technique and some results of the impulsive differential inequality,oscillatory properties of the solutions of certain nonlinear impulsive delay hyperbolic partial differen... In this paper,by making use of the calculous technique and some results of the impulsive differential inequality,oscillatory properties of the solutions of certain nonlinear impulsive delay hyperbolic partial differential equations with nonlinear diffusion coefficient are investigated.Sufficient conditions for oscillations of such equations are obtained. 展开更多
关键词 NONLINEAR IMPULSE DELAY hyperbolic partial differential equations OSCILLATION
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Inhibition of latrunculin-A on dexamethasone-induced fibronectin production in cultured human trabecular meshwork cells 被引量:2
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作者 Yun Wang, Su-Ping Cai 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2011年第3期239-242,共4页
AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells. METHODS:... AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells. METHODS: HTM cells were cultured to confluent and incubated with 0.4 mu mol/L Dex and/or 0.05 mu mol/L LAT-A. FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy. RESULTS: Dex up-regulated FN production in HTM cells, failed to do so when co-incubated with LAT-A. LAT-A decreased production of FN in cultured HTM cells. CONCLUSION: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma. It has an important prospect as an intraocular pressure-lowering drug. 展开更多
关键词 latrunculin-A DEXAMETHASONE human trabecular meshwork cells FIBRONECTIN
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Synergistic effect of combining sulfate reducing bacteria and zerovalent iron permeable reactive barriers on the treatment of groundwater rich in uranium, sulfate and heavy metals 被引量:1
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作者 Zhengji YI Kaixuan TAN +2 位作者 Zhenxun YU Aili TAN Shiqiang WANG 《Chinese Journal Of Geochemistry》 EI CAS 2006年第B08期125-126,共2页
关键词 硫酸盐 细菌 重金属 地下水 水文化学
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Partition of Unity for a Class of Nonlinear Parabolic Equation on Overlapping Non-Matching Grids 被引量:1
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作者 Qisheng Wang Kang Deng +1 位作者 Zhiguang Xiong Yunqing Huang 《Numerical Mathematics A Journal of Chinese Universities(English Series)》 SCIE 2007年第1期1-13,共13页
A class of nonlinear parabolic equation on a polygonal domain Ω R2 is inves- tigated in this paper. We introduce a finite element method on overlapping non-matching grids for the nonlinear parabolic equation based o... A class of nonlinear parabolic equation on a polygonal domain Ω R2 is inves- tigated in this paper. We introduce a finite element method on overlapping non-matching grids for the nonlinear parabolic equation based on the partition of unity method. We give the construction and convergence analysis for the semi-discrete and the fully discrete finite element methods. Moreover, we prove that the error of the discrete variational problem has good approximation properties. Our results are valid for any spatial dimensions. A numerical example to illustrate the theoretical results is also given. 展开更多
关键词 抛物线方程 有限元分析 多角形域 匹配 偏微分方程
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Isolation and bioinformatics analysis of differentially methylated genomic fragments in human gastric cancer 被引量:1
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作者 Ai-Jun Liao Qi Su +2 位作者 Xun Wang Bin Zeng Wei Shi 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第9期1333-1338,共6页
AIM:To isolate and analyze the DNA sequences which are methylated differentially between gastric cancer and normal gastric mucosa. METHODS: The differentially methylated DNA sequences between gastric cancer and normal... AIM:To isolate and analyze the DNA sequences which are methylated differentially between gastric cancer and normal gastric mucosa. METHODS: The differentially methylated DNA sequences between gastric cancer and normal gastric mucosa were isolated by methylation-sensitive representational difference analysis (MS-RDA). Similarities between the separated fragments and the human genomic DNA were analyzed with Basic Local Alignment Search Tool (BLAST). RESULTS: Three differentially methylated DNA sequences were obtained, two of which have been accepted by GenBank. The accession numbers are AY887106 and AY887107. AY887107 was highly similar to the 11th exon of LOC440683 (98%), 3' end of LOC440887 (99%), and promoter and exon regions of DRD5 (94%). AY887106 was consistent (98%) with a CpG island in ribosomal RNA isolated from colorectal cancer by Minoru Toyota in 1999. CONCLUSION: The methylation degree is different between gastric cancer and normal gastric mucosa. The differentially methylated DNA sequences can be isolated effectively by MS-RDA. 展开更多
关键词 Gastric cancer DNA methylation Differentia sequences Methylation-sensitive representationa difference analysis
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Periodic Bifurcation and Soliton Deflexion for Kadomtsev-Petviashvili Equation 被引量:1
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作者 戴正德 李绍林 +1 位作者 李栋龙 朱爱军 《Chinese Physics Letters》 SCIE CAS CSCD 2007年第6期1429-1432,共4页
The spatial-temporal bifurcation for Kadomtsev-Petviashvili (KP) equations is considered. Exact two-soliton solution and doubly periodic solution to the KP-I equation, and two classes of periodic soliton solutions i... The spatial-temporal bifurcation for Kadomtsev-Petviashvili (KP) equations is considered. Exact two-soliton solution and doubly periodic solution to the KP-I equation, and two classes of periodic soliton solutions in different directions to KP-Ⅱ are obtained using the bilinear form, homoclinic test technique and temporal and 1 spatial transformation method, respectively. The equilibrium solution uo =-1/6, a unique spatial-temporal bifurcation which is periodic bifurcation for KP-I and deflexion of soliton for KP-Ⅱ, is investigated. 展开更多
关键词 BOUSSINESQ EQUATION RATIONAL SOLUTIONS EVEN CONSTRAINT HIERARCHY
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Construction of the Eukaryotic Expression Vector for Outer Membrane Protein Tp92 from Treponema pallidum and Its Preliminary Study on the Immune Responses in New Zealand Rabbits 被引量:7
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作者 赵飞骏 吴移谋 +1 位作者 刘双全 余敏君 《Journal of Microbiology and Immunology》 2004年第3期191-196,共6页
To construct the recombinant plasmid of eukaryotic expression containing Tp92 gene from Treponema pallidum and study its immunogenicity in New Zealand white rabbits. Tp92 gene was amplified from the genomic DNA of T. ... To construct the recombinant plasmid of eukaryotic expression containing Tp92 gene from Treponema pallidum and study its immunogenicity in New Zealand white rabbits. Tp92 gene was amplified from the genomic DNA of T. pallidum by polymerase chain reaction (PCR) and subcloned into appropriate site of pcDNA3.1(+) vector. After identification by sequencing and restrictive enzyme digestion, the recombinant plasmid was transfected into HeLa cells using liposome, and the expressed protein was identified by immunocytochemistry and Western blotting. After verifying that the Tp92 antigen gene fragment could be expressed in HeLa cells, 100?μg of recombinant plasmids [pcDNA3.1(+)-Tp92], 100 μg of control plasmids [pcDNA3.1(+)] or 0.5 ml PBS buffer were administered in 3 groups of New Zealand white rabbits (6 rabbits/group), and the booster immunizations were employed at 2-week interval for 3 times. ELISA assay was used for the quantitative detection of the specific antibody in the sera of rabbits, and the proliferation response of spleen cells was detected by MTT assay. It was found that the target gene Tp92 segment about 2103 bp was obtained, and the DNA sequence of Tp92 gene constructed in pcDNA3.1 (+) vector was consistent with the published nucleotide sequence. The homologies of the nucleotide and putative amino acid sequences of Tp92 gene between T.pallidum subsp. pallidum Nichols and various pathogenic treponeme strains were 95.5%-100%. The analysis of immunocytochemistry and Western blotting showed that Tp92 gene segment constructed in pcDNA3.1(+) vector could express a fusion protein with a calculated molecular mass of 77 kDa in HeLa cells and the expressed protein could react with positive blood serum from syphilis patient. The specific antibody IgG titers were observed and the highest titer was 1∶1024 in rabbits after 3 times with pcDNA3.1(+)-Tp92. The proliferation response of spleen cells were significantly higher than that of rabbits injected with pcDNA3.1(+) ( P <0.05). The successful expression of the eukaryotic expression plasmid of Tp92 gene from T. pallidum was obtained in eukaryotic system and strong responses of humoral and cellular immunity was evoked by DNA vaccine of pcDNA3.1(+)-Tp92 in rabbits thus establishing a solid basis for the future studies in the biological activities and for the development of the syphilis DNA vaccine. 展开更多
关键词 Treponema pallidum DNA vaccine Tp92 gene Immune response
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