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1.
TheMelanocyte(MC)isatypeofdendriticcellswhichsynthesizesandsecretesmelanin〔1,2〕.Skinmelaninactsasabarriertothesunlightandultravioletray.InordertoexplorethedevelopmentofMCinphylogeneticprocessandtomakeafurtherunderstandingonthefunctionofMCanddiseasesr…  相似文献   

2.
In1985,DerkneckmadethecloneofTGFβ1fromthegenebankofhuman'splacentaandcDNAoffibromasarcomacellline.PreviousstudieshaveshowedthatTGFβ1existsinmultiplenormalsomaticcells,hematopoieticcellsandembryoniccellsofhumanbeingorothermammals.Cellsandtumortissuewh…  相似文献   

3.
Mesenchymalstemcells(MSCs)weremultipo tentialcells ,locatedinbonemarrow ,skeletalmus cleandsynovialmembrane ,whichcoulddifferenti ateintoseveraltypesofcells,suchasosteocyte ,chondrocyteandadipocyte[1,2 ] .SeperationofMSCsfrombonemarrowwasrelativelyconveniet ,aut…  相似文献   

4.
Keshandisease (KD) ,anendemiccardiomyo pathy ,andKashin Beckdisease (KBD) ,anendemicdeformativeosteoarthropathy ,aretwoindependentdiseaseswithunknowncause .PreviousstudieshaveindicatedthatthebloodSecontentandGSH Pxac tivityofresidentsintheendemicareasofbothdis…  相似文献   

5.
Thetheoriesofchannelsandcollateralsarethecoreofthetraditionalchinesemedicalscience.Thedevelopmentofmodernbiology,especiallythedeepunderstandingofthestructureandfunctionofneuralsystem,provideusstrongevidenceandmethodsinresearchofchannelsandcollaterals…  相似文献   

6.
Endothelialcells (ECs)functionasthebarrierofthebrainandrelatetothesecretionofsomeendo crines.Theirdysfunctioniscloselyrelatedtotheoccurrenceanddevelopmentofcerebralvasculardiseases[1] .DamagetoECsisthefirstsignandaba sicfactorofcerebralischemia .Therefore ,…  相似文献   

7.
CD4 4v6 proteinisacelladhesionmoleculeex pressingonthecellsurface .Recently ,manyscholarsnoticedtherelationbetweenCD4 4v6expressionandneoplasmdevelopmentaswellasprogress ,butthere portsaboutthatarescarceinGBC .InordertostudytheclinicsignificancesofCD4 4v6expres…  相似文献   

8.
IntroductionAsynchronousTransferModehasbeengivenconsiderableattentionasapowerwayofaccom-plishinganintegratedservicenetwork[1].InATMNetworks,informationisdividedintoshortfixed-lengthentities,calledcells,whichareprovidedwithflowidentificationlabels,and…  相似文献   

9.
TelomeraseisanRNAdependentpolymerasethatsynthesizestrlomereTTAGGGrepeatssequencesattheendsofchromosomesrequiredfortheintegrityandstabilityoflinearchromosomes[1] .Normalsomaticcells ,generallylackingtelomeraseactivity ,displayprogressiveattritionoftelomer…  相似文献   

10.
Duringrenalischemia reperfusioninjruy ,nu merousfreeradicalsareproducedandMDA ,aprod uctoffreeradical,andET 1inkidneysbothincreaseintheprocessofreperfusion .SODiscableofelimi natingfreeradicals .AdministrationofSOD priortoischemiaoratthebeginningofreperfusio…  相似文献   

11.
目的 从外周血单核细胞中克隆人IL 1 8成熟编码序列cDNA ,对其进行测序并构建原核表达载体 ,进行原核表达。方法 从人外周血单核细胞中分离总RNA ,进行RT PCR获得人IL 1 8cDNA。测序证实其结果正确后 ,构建原核表达载体。E .coli的表达产物通过SDS PAGE、WesternBlot证实。结果 所克隆的人IL 1 8cDNA编码序列经测序证实与GenBank所报道的序列完全一致 ,细菌表达产物经SDS PAGE证实其大小约为 1 8.3KDa,WesternBlot进一步证实了所表达产物的正确性。结论 本研究结果为进一步探讨IL 1 8的生物学活性和特性奠定了基础 ,同时亦为利用人IL 1 8进行免疫基因治疗的研究打下了良好的基础。  相似文献   

12.
目的 构建HPV 6 / 11L1/E6、HPV 6 / 11L1/E7预防、治疗性DNA疫苗质粒。方法 运用PCR扩增HPV 6 /11L1、E6、E7序列 ,PCR产物连接至pGEMT Easy质粒 ,测序鉴定正确后 ,分别连接至真核表达载体pVAX ,再用酶切、连接而构建成HPV 6 / 11L1 E6 /E7 pVAX质粒。运用电穿孔法将所获得质粒转染COS 7细胞 ,免疫组化检测蛋白表达情况。结果 所构建质粒的插入目的片段测序正确 ;免疫组化检测在胞浆胞核可见棕黄色点状阳性产物沉积。结论 所构建的HPV 6 / 11L1 E6 /E7 pVAX融合蛋白表达质粒可在体外表达L1 E6 /L1 E7蛋白 ,为今后进行DNA疫苗的动物实验及临床实验研究做好准备  相似文献   

13.
Objective To construct eukaryutic expression vector of HPV18 L1- E6, E7 chimeric gene and examine the humorul and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major rapsid protein L1 gene and mutant zinc finger domains of early E6/7 oncogenes in HPV18 were integrated and inserted into eukaryotic expression vector pVAX1 to generate vaccines pVAX1-L1E6Mxx, ETMxx. CHO cells were transiently transfected with the individual construct. Target protein expressions in the lysate of the transfected cells were measured by ELISA and immunocytochemistry After BALB/c mice were vaccinated with various recombinant plusmids(pVAX1- L1-E6M3 or pVAX1-L1-E7M3 ) and immunie adjuvants (pLXHDmB7-2 or LTB) through different administration routes (intramuscular or intranasal) , the great cellular immune responses were produced us revealed by delayed-type hypersensitivity (DTH) and lymphocyte proliferation, and the expression of IL-4 and IFN- 7 cells in CD4^+ and CD8^+ subpopulations. Results The highly efficient expression of pVAX1-L1E6Mxx, E7Mxx vector in host eukaryotic cells were demonstrated both by ELISA and immunocytochemistry. The level of specific serum IgG against HPV in experiment groups mice was much higher than that of control group, and intranuscular immunization group had the highest antibody level. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8^+ IFN-γ^+ cells number, but CD4^+ IL4^+ cell number was higher in intranasul immunization groups. The immunization groups using pLXHDmB7.-2 as adjuvant were superior to other groups in immunorespouse. Conclusion These DNA vaccines produce remarkable cellular and humorul immune responses in the mouse and may provide us prophylatic and therapeutic candidates for HPV induced cancer treatment.  相似文献   

14.
克隆人 IL - 1 2 ( h IL - 1 2 ) p40和 p35亚单位 c DNA,构建人单链 IL- 1 2 ( rhsc IL- 1 2 )融合基因 ,并在哺乳动物细胞中进行表达。方法 从经 PDBu刺激的 EBV转化的人 B淋巴母细胞株 NC37中提取 m RNA,经 RT- PCR分别获得 h IL- 1 2 p40和 p35亚单位编码序列的 c DNA,运用重组 PCR技术将两段基因通过一疏水性多肽接头 ( Gly4 Ser) 3 DNA序列进行体外基因重组 ,构建 rhsc IL- 1 2融合基因 ,将其插入 pc DNA3.1 ( + )真核表达载体 ,经脂质体转染 COS7细胞进行表达 ,Western blot进行分析。结果 所克隆的 h IL- 1 2 p40、p35c DNA序列和构建的 rhsc IL - 1 2融合基因 DNA序列均经测序得以证实 ,融合基因可在 COS7中表达其产物 rhsc IL- 1 2融合蛋白 ,其分子量为 70 KD,可与鼠抗人 IL - 1 2 p40 /p70单克隆抗体特异性结合。结论 本研究结果为进一步探讨 rhsc IL- 1 2融合蛋白的生物学活性和特性奠定了基础 ,也为 rhsc IL- 1 2融合基因在原核细胞中的表达提供了可能性  相似文献   

15.
为了构建pcDNA-IL-2真核表达质粒,并探讨白细胞介素-2(IL-2)cDNA导入对顺铂诱导的卵巢癌耐药细胞株耐药性的影响。我们采用PCR技术扩增人IL-2cDNA片段,构建pcDNA-IL-2真核表达质粒。导入顺铂诱导的人卵巢癌耐药细胞株,观察其耐药性的改变。结果成功构建了pcDNA-IL-2真核表达质粒,并在Cos-7细胞高效表达,导入卵巢癌耐药细胞后其对顺铂的敏感性显著升高。说明转导pcD-NA-IL-2基因的人卵巢癌耐药细胞株其耐药性有明显逆转,而且IL-2亦可逆转非mdr1依赖的耐药性  相似文献   

16.
Herpessimplexvirustype 2 (HSV 2 ) ,onekindofdoublestrandsDNAvirus ,cancauseseriousworldwildinfections.Herpessimplexvirusgenom icDNAcodesforapproximately 80 proteins .Mostorallareexpressedduringlyticinfection ,manyarefoundinintactvirionsandatleast 11havebeenident…  相似文献   

17.
放射诱导p16基因胰腺癌靶向性表达的研究   总被引:1,自引:0,他引:1  
目的 构建 pcDNA3.1 Egr.1p-p16重组质粒并检测其在人胰腺癌 JF305细胞中的辐射诱导表达。方法 将人p16 cDNA基因连接到有辐射诱导特性的早期反应因子 Egr.1p的下游,构建成 pcDNA3.1 -Egr.1p- p16 重组质粒,利用脂质体介导转染人胰腺癌细胞系 JF305 细胞;采用 RT- PCR方法和 Western blot方法检测不同剂量 X射线照射后,被转染细胞中 p16 的转录和表达水平。结果 酶切鉴定证实 pcDNA3. 1 Egr. 1p -p16 重组质粒构建正确。被pcDNA3.1 Egr.1p -p16重组质粒转染的人胰腺癌 JF305细胞经不同剂量 X射线照射后,p16 基因表达均高于未照射组。结论 X射线可诱导 pcDNA3.1 -Egr.1p -p16重组质粒在人胰腺癌 JF305细胞中表达增强。  相似文献   

18.
目的克隆并构建乙型肝炎病毒(HBV)全X基因表达载体,探讨HBV全X基因对HEK293细胞功能的影响,并比较HBV全X基因与X基因对细胞影响的差异。方法应用基因克隆的方法从慢性HBV感染者血清中获得HBV全X及X基因,构建带有HBV全X基因与X基因的真核表达载体以及带有绿色荧光基团GFP的重组质粒;细胞转染技术将其分别转入HEK293细胞,RT-PCR及Western-blot技术验证HBV全X基因及X基因在细胞内的表达;荧光显微镜下观察其在细胞内的定位情况;MTT法检测细胞增殖率的变化;Annexin V流式细胞分析技术检测其对细胞凋亡率的影响;TRAP实时荧光定量PCR法检测各组细胞端粒酶活性的变化。结果成功构建带有HBV全X基因的重组质粒且目的基因能在HEK293细胞内高表达;表达的HBV全X蛋白主要分布在胞核,而X蛋白在胞质、胞核均有分布;转染HBV全X和X基因后细胞增殖率分别为(0.826±0.002)%和(0.805±0.013)%,显著高于转染空质粒(0.691±0.035)%和未转染对照组(0.681±0.018)%,差异具有统计学意义(P<0.05);转染HBV全X和X基因后细胞凋亡率分别为(25.25±3.02)%及(21.34±2.16)%,显著高于转染空质粒(13.49±1.49)%及未转染细胞对照组(11.37±1.58)%,差异具有统计学意义(P<0.05);转染全X及X基因细胞的端粒酶活性亦显著高于对照组,差异有统计学意义(P<0.05);全X转染组细胞的增殖率、凋亡率及端粒酶活性均高于X转染组,但差异均无统计学意义(P>0.05)。结论 HBV全X基因具有促细胞增殖、凋亡以及影响细胞端粒酶活性的作用,其可能通过不同于HBV X基因的细胞内途径影响细胞的生物学功能,从而在HBV相关的原发性肝细胞癌发生发展中起作用。  相似文献   

19.
目的 构建带有 6×myc的人白介素 17受体样分子的重组表达质粒 ,以便检测hIL 17RLM L基因在真核细胞中的特异性表达。方法 设计带有酶切位点 (EcoRⅠ和XhoⅠ )的特异性引物 ,用PCR方法扩增hIL 17RLM L片段回收后插入带有 6×myc标签的 pcDNA3.0真核表达载体 ,转染COS7细胞后作Westernblot检测其表达。结果 成功地构建了带有 6×myc标签的 pcDNA3.0 6×myc /hIL 17RLM L重组质粒 ,Westernblot检测到该质粒可在真核细胞中表达。结论 用分子生物学的方法成功地构建了真核表达质粒pcDNA3.0 6×myc /hIL 17RLM L ,使该基因的特异性检测成为可能 ,为进一步研究hIL 17RLM L的生物学功能奠定了基础。  相似文献   

20.
Cervicalcancerisacommonmalignancyofmarriedwomen.HighriskHPV(HPV16,18)infectioniscloselyassociatedwithcervicalcancer(l).HPV16DNAcanbedetectedinapproximately50%~90?rvicalcarcinomastZ).HPV16E6andE7geneshasbeenprovedtobetransforminggenes.TheirgeneproductscanbindtoandinactivateP53andPRbrespectively,playingacausalroleinthedevelopmentofcervicalcarcinomat27.AlotofinvestigationindicatethatHPV16E6/E7wereconsistentlyandhighlyexpressedincervicalcarcinomas,andthatE6/E7proteinscouldinducebothhu…  相似文献   

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