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31.
抗人大肠癌单链抗体的构建、表达及其体内外生物学活性的检测 总被引:1,自引:2,他引:1
目的 将抗人大肠癌单克隆抗体ND-1(mAb)的VR和VL基因进行重组,构建和表达ND-1scFv,并对其在体内外的生物学活性进行检测。采用RT-PCR技术,从能够分泌mAb ND-1的鼠杂交瘤细胞中扩增VH和RL基因,通过重叠延伸拼接PCR在VH和VL基因间引入连接短肽,体外构构建ND-1scFv基因,并在大肠杆菌中表达。采用间接免疫荧光(IFA)EY IF工IMPG-1scFv的免疫学活性。用^99Tc^m标记ND-1scFv后,将偶联物给予荷瘤裸鼠,观察其在动物体内的显像及生物学分布。结果 SDS-PAGEW显示,重组蛋白Mr为30000,同预期结果一致。IFA及ELISA检测表明,ND-1scFv保留了与亲本抗体相近的免疫学活性,对表达相应抗原的靶细胞具有行异结合活性。体内放射免疫实验显示,^99Tc^m-ND-1scFv在荷瘤小鼠体内的生物学分布,呈明显的肿瘤积聚趋向,注入体内1h血中T/NT即在2.61。结论 获得免疫学活性良好的ND-1scFv,对荷瘤动物体内肿瘤的定位快速,准确,可望成为有效的肿瘤诊断和治疗的导向载体。 相似文献
32.
33.
A 63-year-old man with iron loss anaemia and hypercalcaemia was found to have a renal cell carcinoma. Despite the iron-deficient blood and bone marrow picture, the serum ferritin concentration was markedly raised. This was mainly due to a “basic isoferritin”. The serum parathormone concentration was normal. The serum ferritin and calcium concentrations returned to normal after the tumour was removed. We propose that the renal cell carcinoma cells in this patient secreted the basic isoferritin as well as humoral factor(s) responsible for hypercalcaemia. 相似文献
34.
体外厌氧条件下载铜蒙脱石杀菌效果的研究 总被引:9,自引:0,他引:9
载铜蒙脱石的杀菌作用通过脑心浸液肉汤 (BHI)二倍系列稀释法来判定其最小抑菌浓度 (MIC)。释放入肉汤和生理盐水中Cu2 的量通过原子吸收光谱仪测定。杀菌动力学研究采用改良的振荡瓶法。结果表明 :厌氧条件下 ,载铜蒙脱石具有很强的杀菌性能。载铜蒙脱石对放线共生放线杆菌的最小抑菌浓度为 2 .5 6mg/ml;对血液链球菌为 5 .12mg/ml。测得肉汤二倍稀释的载铜蒙脱石释放出的Cu2 量在 2 .39~ 38.6 5 ppm之间 ,在生理盐水中释放的Cu2 量为 1.85~ 15 .82ppm。本试验结果表明 ,蒙脱石无抗菌性能。 相似文献
35.
SARS疫情期间公众心态影响及变化趋势分析 总被引:9,自引:0,他引:9
目的:探索重大突发性公共卫生事件对社会公众心理行为的影响,以及在不同阶段中公众心态变化的特点。方法:对SAILS流行期间374人次的心理咨询记录进行分析,依据疫情发展情况,比较各阶段的差异。结果:在咨询者中,女性(59.4%)多于男性(40.6%);根据疫情变化将咨询记录数据划分为三个阶段,分析发现随着疫情缓和,针对SARS症状的咨询者显著减少,从13.1%降低到1.8%;有关社会生活的咨询者显著增多,从8.7%上升到12.3%;在有关情绪问题的咨询者中,伴有相关症状者显著减少,从23.0%降低到4.5%;不伴相关症状者显著增多,从77.0%上升到95.5%。结论:整个SAILS疫情期问,情绪问题始终占第一位;随SARS疫情缓和,社会公众对于疾病本身的关注减少,而与疫情有关的社会生活问题增多;面对突发性公共卫生事件,及时开通心理咨询热线电话是十分必要的。 相似文献
36.
骨髓源间充质干细胞在异基因小鼠免疫器官内的分布及其免疫功能调节作用 总被引:20,自引:2,他引:20
目的:探讨骨髓源间充质干细胞(Bone marrow mesenchymal stem cells,BMSC)在异基因小鼠免疫器官内的分布及其免疫调节作用。方法:以CM-Dil荧光染料示踪BMSC的体内分布情况,并辅以PCR检测Y染色体的方法进一步鉴定;体外实验采用MTT法、ELISA和FACS等方法检测BMSC的免疫调节作用。结果:BMSC可进入并较长期(30天)存在于异基因小鼠免疫器官内;在体外,BALB/C小鼠的BMSC对由ConA诱导的BALB/C和C57BL/6(B6)和BXSB小鼠的T细胞增殖均有抑制作用;而对前两种小鼠由12S诱导的B细胞增殖和分泌k方面表现为促进作用,对BXSB小鼠由IPS诱导的B细胞增殖和k分泌有抑制作用。BALB/C小鼠的BMSC对BALB/C和B6小鼠由ConA诱导的IL-4生成细胞的数量无明显影响,却可降低由ConA诱导的两种品系小鼠的IFN-γ生成细胞的数量;但对于BXSB小鼠却不同,BALB/C的BMSC可降低由ConA诱导的BXSB小鼠的IL-4生成细胞的数量,而提高由ConA诱导的IFN-γ生成细胞的数量。结论:异基因BMSC不但可进入受体的免疫器官,且可较长期(30天)存在;另外,BMSC对同基因正常、异基因正常和异基因自身免疫病的个体均有一定程度的免疫调节作用。 相似文献
37.
流感病毒血凝素基因工程抗体的抗病毒实验效果观察 总被引:1,自引:0,他引:1
目的:对昆虫细胞系统表达纯化的中和性流感病毒血凝素基因工程全抗体IgG-IV-2,IgG-IV-6进行体外和体内抗病毒效果的研究。方法:对比抗体应用前后病毒在MDCK细胞中的病毒滴度和动物模型中粘膜用药前后鼠肺内病毒滴度的改变,验证抗体的粘膜抗感染效果。结果:两株基因工程抗体使4.5log10TCID50滴度的病毒下降1/2的剂量分别为0.8μg和0.5μg;动物模型的粘膜给药表明使4.0log10TCID50的病毒下降1/2所需的抗体剂量IgG-IV-2为0.25mg/kg体重,IgG-IV-6为0.1mg/kg体重,联合应用时为0.08mg/kg体重。结论:获得的基因工程抗体具有体内体外的抗病毒效果,能够中和病毒毒力。 相似文献
38.
Gene conversion is a likely cause of mutation in PKD1 总被引:3,自引:0,他引:3
Watnick TJ; Gandolph MA; Weber H; Neumann HP; Germino GG 《Human molecular genetics》1998,7(8):1239-1243
Approximately 70% of the gene responsible for the most common form of
autosomal dominant polycystic kidney disease ( PKD1 ) is replicated in
several highly homologous copies located more proximally on chromosome 16.
We recently have described a novel technique for mutation detection in the
duplicated region of PKD1 that circumvents the difficulties posed by these
homologs. We have used this method to identify two patients with a nearly
identical cluster of base pair substitutions in exon 23. Since pseudogenes
are known to be reservoirs for mutation via gene conversion events for a
number of other diseases, we decided to test whether these sequence
differences in PKD1 could have arisen as a result of this mechanism. Using
changes in restriction digest patterns, we were able to show that these
sequence substitutions are also present in N23HA, a rodent-human somatic
cell hybrid that contains only the PKD1 homologs. Moreover, these changes
were also detected in total DNA from several affected and unaffected
individuals that did not harbor this mutation in their PKD1 gene copy. This
is the first example of gene conversion in PKD1 , and our findings
highlight the importance of using gene-specific reagents in defining PKD1
mutations.
相似文献
39.
Alexandra P. Key Hatun Zengin‐Bolatkale Anastasia Dimitropoulos Ellen Doernberg 《American journal of medical genetics. Part A》2020,182(7):1655-1663
This study examined sensitivity of eye tracking measures to hyperphagia severity in Prader‐Willi syndrome (PWS). Gaze data were collected in 57 children with PWS, age 3–11 years, and 47 typically developing peers at two study sites during free visual exploration of complex stimulus arrays that included images of food, animals, and household objects. Analysis of the number and duration of fixations as well as gaze perseverations revealed that food items are not exceptionally salient for children with PWS. Instead, increased attention to food in the context of other high‐interest items (e.g., animals) was associated with caregiver reports of more severe hyperphagia and more advanced nutritional phase. The study also provided preliminary evidence of possible genetic subtype and sex differences as well as demonstrated that multiple investigators in a wide range of settings can effectively implement the eye tracking protocol. The results indicate that gaze characteristics derived from eye tracking may be a promising objective marker of hyperphagia in PWS for use in research and clinical trials. 相似文献
40.
Predominance of null mutations in ataxia-telangiectasia 总被引:15,自引:4,他引:15
Gilad S; Khosravi R; Shkedy D; Uziel T; Ziv Y; Savitsky K; Rotman G; Smith S; Chessa L; Jorgensen TJ; Harnik R; Frydman M; Sanal O; Portnoi S; Goldwicz Z; Jaspers NG; Gatti RA; Lenoir G; Lavin MF; Tatsumi K; Wegner RD; Shiloh Y; Bar-Shira A 《Human molecular genetics》1996,5(4):433-439
Ataxia-telangiectasia (A-T) is an autosomal recessive disorder involving
cerebellar degeneration, immunodeficiency, chromosomal instability,
radiosensitivity and cancer predisposition. The responsible gene, ATM, was
recently identified by positional cloning and found to encode a putative
350 kDa protein with a Pl 3-kinase-like domain, presumably involved in
mediating cell cycle arrest in response to radiation-induced DNA damage.
The nature and location of A-T mutations should provide insight into the
function of the ATM protein and the molecular basis of this pleiotropic
disease. Of 44 A-T mutations identified by us to date, 39 (89%) are
expected to inactivate the ATM protein by truncating it, by abolishing
correct initiation or termination of translation, or by deleting large
segments. Additional mutations are four smaller in-frame deletions and
insertions, and one substitution of a highly conserved amino acid at the Pl
3-kinase domain. The emerging profile of mutations causing A-T is thus
dominated by those expected to completely inactivate the ATM protein. ATM
mutations with milder effects may result in phenotypes related, but not
identical, to A-T.
相似文献