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1.
Leishmaniaparasiteinfectionnotonlycausescateoneousandmucosallesions,butalsoviseralLeishmaniasis1 ThestudyindicatedthatLeishmaniainfectioncouldevokeinfectedsubject’simmunesystemandacquiredprotectiveimmunityandinhibitsecondaryLeishmaniainfections2 So…  相似文献   

2.
Thetraceelementselenium (Se)servesastheactivecenterofcytosolicglutathioneperoxidase (GPX 1)intheformofSe cysteine GPX 1isanimportantenzymeinthecellantioxidizingdefensesystem ,scavenginghydroxylperoxidesandlipidhydroperoxidesinlivingcells 1 TheabundanceofGPX 1…  相似文献   

3.
Wilms’tumor(WT1)geneisatumorsupressorgeneidentifiedinWilms’tumorpatientsItislocatedonchromosome11p131 ThelengthofWT1geneisabout50KbThereare2splicingexonsoutofits10exons:exon5(spliceⅠ)andexon9(spliceⅡ)Thepresenceoftwoalternativesplicesresultsinfo…  相似文献   

4.
Hepatocellularcarcinoma (HCC)ranksthirdamongthecausesofcancermortalityinChina ,andabout4 2 5%oftheworld’snewcasesofHCCeachyearoccurinChina 1EpidemiologicstudiesprovideevidencethatinfectionwithhepatitisB (HBV )and/orhepatitisC (HCV ) ,andingestionofaflatoxinBcon…  相似文献   

5.
ObjectiveTosearchdiferentialyexpressedsequencescorelatedwithpathogenesisofhumannasopharyngealcarcinoma(NPC),includingthecandi...  相似文献   

6.
Theecotropicvirusintegrationsite1(Evi1)gene,locatedatchromosomeband3q26,wasoriginallyidentifiedasageneassociatedwithretrovirallyinducedmyeloidleukemiainmice1,2 ManystudieshaveshownthattheEvi1geneisoverexpressedinmyeloidleukemiawith3q26,involvingchromosomalab…  相似文献   

7.
InChina,hepatocellularcarcinoma(HCC)rankssecondincancermortalitysince1990s-InthefieldofHCCtreatment,surgicalresectionremainsthebest,whichincludeslargeHCCresection,smallHCCresection,reresectionofsubclinicalrecurrence,aswellascytoreductionandsequentia…  相似文献   

8.
Itissuggestedthatexcessfreeradicalformationmayoccurinpatientswithschizophrenia14 Whenthenumberoffreeradicalincreases,theenzymesinvolvedinradicalmetabolismmaybeconsequentlyalteredSuperoxidedismutase(SOD)isoneofthescavengingenzymesdetoxifyingsuperoxidera…  相似文献   

9.
Chronicperiodontitisisaverycommonoraldisease Sinceitisadifficultdiseasetocure ,itattractstheattentionofmanyresearchers Recentstudiessuggestthatimmunocompetencesubstancespresentinthehumanbodyareimportantfactorsinphysiologicalfunctionanddemonstratethatel…  相似文献   

10.
SchistosomiasisisoneofthemostseriousparasiticdiseasesintheworldInviewoftheimportanceofegggranulomasinthepathogenesisofschistosomiasis,clearunderstandingofthemechanismsofgranulomaformationandmodulationisthereforenecessaryforthecontrolofthediseaseInt…  相似文献   

11.
联合运用四种技术进行血友病A的基因诊断   总被引:20,自引:0,他引:20  
目的:最大限度地提高血友病A(HA)患者及家系成员的基因诊断、携带者检出及产前诊断的可诊断率。方法:对于26例HA患者和家系的女性家属首先采用长距离DNA扩增(LD-PCR)技术,直接检测是否为FⅧ基因倒位及其携带者;对于非倒位的HA家系依次采用Bcl I PCR/RFLP分析技术、基因内含子13(CA)n、内含子22(GT)n(AG)n二核苷酸重复序列多态性分析技术以及与FⅧ基因紧密连锁的可变串联重复序列多态性分析技术(St14 VNTR/PCR)进行间接诊断。结果:在26个HA家系的16个重型家系中查出7个基因倒位,占重型HA的43.8%。19个非倒位HA家系,用上述三种间接诊断技术分别有16、13及17个HA家系可以作出诊断,可诊断率分别为84.2%、68.4%和89.5%,联合上述四种技术,对26个HA家系全部作出了诊断。结论:联合采用四种基因诊断技术,几乎可以为所有有家庭史的HA家系作出基因诊断及携带者检出。  相似文献   

12.
LD-PCR直接基因诊断重型血友病A的研究   总被引:1,自引:1,他引:0  
目的:检测凝血因子VⅢ基因倒位,提高对重型血友病A患者及其携带者的诊断水平。方法:根据患者的出血症状及遗传史,采用国际通用的一期法检测患者血浆凝因因子VⅢ活性(FVⅢ:C),ELISA法检测血浆vWF:Ag浓度,确诊血友病A及其携带者,对38例重型HA患者及其母亲或姨母,采用长距离DNA扩增(LD-PCR)技术检测是否存在凝血因子VⅢ(FVⅢ)基因倒位,进行直接基因诊断。结果:38例无亲缘关系的重型血友病A患者中,发现15例患者(或家系)有FVⅢ基因倒位,占重型患者的41%;该15例家系中查出基因倒位携带者5名,结论:利用LD-PCR检测FVⅢ基因倒位技术可以准确,简便而快速地直接进行重型血友病A的基因诊断和携带者检测。  相似文献   

13.
目的 分析山东省55例血友病A(HA)患者基因突变类型,探讨HA发病机制。 方法 采用LD-PCR法检测山东省55例HA患者凝血因子Ⅷ(FⅧ)基因的内含子22倒位突变,阴性者采用高通量测序法检测FⅧ基因的26个外显子,并与人类基因突变数据库比对。 结果 55例HA患者中内含子22倒位突变20例,其他35例共检测到28种突变类型,其中3例同时检测出2种突变,有6例未检测到基因突变。共发现c.2034_2062del、c.6052_6058del、c.6046_6047del、c.1009+1G>T、c.2997dupA、c.4001delT、c.1334T>A、c.1493G>A、c.557A>T等9种未被报道过的新突变。 结论 FⅧ基因突变是导致HA的根本原因。重型血友病患者中最常见的突变类型为内含子22倒位突变。编码区突变最常见的突变类型为错义突变。新发现的9种新突变丰富了基因突变谱,为探讨HA发病机制提供了基础。  相似文献   

14.
目的 建立能同时筛查A型、B型和新型甲型H1N1流感病毒的多重RT-PCR技术.方法 针对A型流感病毒的M基因、B型流感病毒的NS基因设计通用扩增引物,针对新型甲型H1N1流感病毒的HA基因设计特异性扩增引物,采用一步法建立多重RT-PCR反应体系.通过盲法实验与实时荧光RT-PCR进行比对来评价方法的准确性,并应用于临床评价方法的实用性和有效性.结果 琼脂糖凝胶电泳分析多重RT-PCR产物,结果显示目的扩增片段条带清晰明亮,没有非特异性产物出现,可见该方法扩增效率高,特异性强.50份样本的盲法实验结果显示两种方法检测结果完全一致,符合率为100%.结论 建立的多重RT-PCR方法能通过一次实验快速、准确地同时筛检A型、B型和新型甲型H1N1流感病毒,是一项成本低廉、对流感的疫情监测和早期诊断具有实用价值的筛检技术.
Abstract:
Objective To developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1)influenza viruses. Methods Two pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated. Results The RT-PCR products were analyzed using agarose gel electrophoresis,which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%. Conclusion This multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.  相似文献   

15.
2种PCR方法扩增盐藻肌动蛋白基因3'旁侧序列比较   总被引:4,自引:0,他引:4  
目的:比较扩增盐藻肌动蛋白基因3’旁侧序列的2种PCR方法。方法:用pvuⅡ、EcoR Ⅴ和StuⅠ3种限制性内切酶消化盐藻基因组DNA后,供2种PCR用,一种是连接介导法PCR(LMPCR),与用人工合成的适配子相连并用适配子引物和盐藻肌动蛋白基因特异引物扩增未知序列;另一种是反向PCR(IPCR),酶切后的DNA自身环化做模板,用2对基因特异引物反向扩增。结果:2轮PCR后,LMPCR中得到大量的非特异扩增产物,测序结果发现许多产物是由适配子引物AP2单独扩增引起。而反向PCR在得到pvuⅡ和Stu Ⅰ消化的自连文库中扩增得到2.5kb特异产物,经测序发现,片段两侧为基因特异引物,部分序列与已知序列相一致。Southern也进一步证实了IPCR扩增片段来源于盐藻基因组DNA。结论:IPCR技术在克隆基因旁侧序列时优于LMPCR方法。  相似文献   

16.
目的 检测中国血友病A(HA)患者中凝血因子Ⅷ(FⅧ)基因内含子1倒位(inv1)的发生频率,并和国外相关资料相比较,明确部分中国HA患者的发病机制.方法 用一期法检测158例无关家系HA患者的FⅧ活性(FⅧ:C),进行HA表型诊断;分别采用长距离和双管多重PCR技术检测内含子22倒位(inv22)和invl;直接测序法进行FⅧ基因全长序列分析.结果 在158例无关家系的HA患者中发现有2例(家系)invl阳性,检出率为1.26%;对其中1例阳性患者家系进行调查,发现1例罕见女性HA患者为invl携带者.对女性患者另一条染色体FⅧ基因进行全长测序,未发现有新基因突变.结论 invl在中国HA人群中发生率相对较低.女性HA患者为inv1杂合子,其发病考虑为与X染色体非随机失活有关.  相似文献   

17.
输血传播病毒(TTV)基因变异的临床意义   总被引:2,自引:0,他引:2  
目的 :研究TTV基因变异与其致病性的关系。方法 :采用巢式多聚酶链式反应 (PCR)扩增TTVDNA ,收集TTVDNA阳性病例 5 2例 ,其中献血员 4例 ,乙型肝炎病毒携带者 5例 ,慢性乙型肝炎患者 11例 ,慢性重型乙型肝炎患者 32例 ,采用单链构象多态性 (SSCP)方法进行TTV基因变异检测。结果 :献血员、乙型肝炎病毒携带者、慢性乙型肝炎、慢性重型乙型肝炎患者中 ,SSCP电泳条带数分别为 1.2 5± 0 .5 0 ,1.6 0± 0 .5 5 ,3.36± 1.36 ,4 .5 9± 1.83;慢性乙型肝炎和慢性重型乙型肝炎患者组中SSCP电泳条带数显著多于其他组 (P <0 .0 5 ) ;慢性重型乙型肝炎患者中SSCP电泳条带数显著多于慢性乙型肝炎患者 (P <0 .0 5 )。结论 :TTV存在基因变异 ,TTV基因变异株的复杂性或称准种感染的复杂性可能是TTV与HBV感染者重叠感染使病情加重的因素之一  相似文献   

18.
输血传播病毒(TTV)基因变异的临床意义   总被引:11,自引:0,他引:11  
OBJECTIVE: To investigate the clinical significance of gene variant of transfusion transmitted virus (TTV). METHODS: A nested polymerase chain reaction (nested-PCR) assay with specific primers from open reading frame one (ORF1) of TTV genome was established to detect TTV DNA in the serum samples from patients with hepatitis B and blood donors. Asymmetric PCR was established to get single strand DNA in the TTV DNA positive cases, and then single strand conformation polymorphism (SSCP) was applied. RESULTS: 1.25 +/- 0.50, 1.60 +/- 0.55, 3.36 +/- 1.36 and 4.59 +/- 1.83 SSCP bands were detected in the blood donors, chronic HBV carriers, chronic hepatitis B patients and chronic severe hepatitis B patients respectively. SSCP bands were much more complicated in chronic hepatitis patients and chronic severe hepatitis than those of chronic HBV carriers and blood donors (P < 0.05). CONCLUSION: There is gene variant in TTV infection. The complication of gene variant strain or quasispecies infection of TTV may be one of the causes responsible for clinical exacerbation of TTV superinfection in chronic hepatitis patients.  相似文献   

19.

Background  Hemophilia A (HA) is an X-linked inherited bleeding disorder caused by decreased activity of factor VIII (FVIII) due to heterogenous mutations in the FVIII coding gene (F8). The type of mutation plays an important role in the FVIII inhibitor formation. To date, several studies on the spectra of F8 defects have been performed in Western populations, but similar studies in Asian races are scarce. Here, we reported the distribution of the F8 gene mutations in 18 unrelated Chinese patients with HA.
Methods  Intron 22 and intron 1 inversions in the F8 gene were screened in 158 unrelated patients with HA using a long-distance PCR and multiplex PCR method. Direct sequencing of the coding region of the F8 gene was used to identify the mutations responsible for HA in 18 unrelated Chinese HA patients who were negative for intron 22 and intron 1 inversions; sequences were compared with the HAMSTeRS database. A clotting method was used to assay the FVIII activity level and the Bethesda assay was used to detect the FVIII inhibitor.
Results  A total of 18 different HA F8 mutations were identified, seven of which were described for the first time. These novel mutations included five small deletions, one point mutation and one small insertion. One novel mutation (4382-3 AC deletion) was associated with inhibitor development.
Conclusion  These data extend our insight into the mechanisms by which novel amino acid mutations may lead to HA and how the HA patient genotypes influence the risk of FVIII inhibitor.

  相似文献   

20.
目的:建立一氧化氮(NO)前体化合物L-精氨酸激活人肝细胞L02内源凝血因子Ⅷ(FⅧ)重表达的分子细胞生物学模型,探讨NO信号激活L02中内源FⅧ重表达的调控通路及分子基础。方法:取对数生长期L02细胞随机分为对照组、加药组(L-精氨酸)、抑制剂组(L-NAME和L-精氨酸)和抑制剂对照组(L-NAME),分别培养12、24、 36、48和60 h,采用流式细胞术检测作用48 h后L02中人FⅧ蛋白的表达, Griess法检测不同时间点L02细胞内NO水平, RT-PCR法检测L02中人FⅧ基因、诱导型一氧化氮合酶(iNOS)基因、核转录因子(NF-κB1)基因和核因子κB(免疫球蛋白κ轻链基因增强子)抑制蛋白α亚基(I-κB alpha)基因的转录水平,Western blotting法检测L02中人磷酸化I-kappaB(磷酸化I-κB)表达水平。结果:流式细胞术检测,加L-精氨酸培养48 h后L02中出现人FⅧ蛋白的表达。Griess法检测,加药组L02在3、6和12 h内NO表达水平显著增加(P<0.05),随后又基本恢复到正常水平,正常组、抑制剂组和抑制剂对照组L02细胞内NO水平无变化;RT-PCR检测,加药组L02中人FⅧ mRNA转录;对照组、抑制剂组和抑制剂对照组均无人FⅧ mRNA的转录,加药组iNOS、NF-κB1和I-κB alpha转录水平均上升(P<0.05),对照组、抑制剂组和抑制剂对照组上述基因转录水平均无明显变化。Western blotting法检测,加入L-精氨酸后,磷酸化I-κB表达水平明显增加(P<0.05),其他组则无此变化。结论:L-精氨酸通过NO信号通路进而激活I-κB磷酸化,导致人FⅧ基因启动子上游调控相关的转录因子NF-κB1进入胞核从而激活人离体肝细胞L02中内源人FⅧ的表达。  相似文献   

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