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1.
用人工合成沙眼衣原体主要外膜蛋白(MOMP)氨基端1/4肽链免疫8周龄雄性BALB/c小鼠3只,第14天用L1/440/Bu原体加强免疫,第24天将免疫反应良好的1只小鼠脾细胞与NS-1瘤细胞融合。用1/4MOMP和L1原体抗原包被的聚丙乙烯板检测上清中抗体,淘汰与鹦鹉热衣原体种(EAE株)、肺炎衣原体种(ATCCVR1310株)以及正常组织培养细胞(BGMK)有交叉反应的克隆,最后得到4株抗沙眼衣原体特异性单克隆抗体(MAbs),其抗体属IsG1和IgG2a亚类。微量免疫荧光试验(micro-IF)发现4株MAbs均与本实验室制备的沙眼衣原体L1、L2、A、B、C、E原体及L2包涵体抗原发生反应,并与沙眼衣原体所有15个标准血清型结合,其腹水MAbs滴度≥1:12800。免疫印迹试验显示4株MAbs均与沙眼衣原体分子量为40000的MOMP反应。  相似文献   

2.
本文以HER-2/neu癌基因表达产物为抗原免疫BALB/C小鼠,采用常规淋巴细胞杂交瘤技术立一株稳定分泌抗P^185McAb的杂交瘤株5E12,McAb5E12属IgG亚类,与SKBR-3,T47D乳癌细胞系呈阳性反应,与HT29,EC109,DAC-1,SP2/0,成纤维细胞及小鼠NIH3T3等细胞系列呈阴性反应,30例乳癌组织石蜡切片同时以自制抗P^185McAb5E12及进口McAbC-e  相似文献   

3.
用丙型肝炎病毒(HCV)核心区合成肽作抗原免疫BALB/c小鼠,取其脾细节与骨髓瘤细胞SP^2/0进行融合。经有限稀释法克隆3代后获得一株抗HCV核心抗原CP10的单克隆抗体(McAb)杂交瘤细胞株3E7,并对其分泌抗体进行初步鉴定。杂交瘤细胞培养上清的抗体滴度为1:320,诱生同系小鼠腹水的滴度为1:12800,该McAb能与CP10特异性结合,杂交瘤细胞染色体数目为106条。分泌的抗体为IgG  相似文献   

4.
以HSV-1为抗原,免疫BALB/C鼠,取其脾细胞在PGE介导下与SP2/0细胞常规融合,IFA法筛选鉴定,反复克隆,共获得7株能稳定、持续分泌抗HSV的McAb的杂交瘤细胞系。核型分析染色体数为87~103条之间,Ig类型分别为LgG_1、IgG_(2g)、IgG_3,腹水抗体效价在10 ̄(-5)~10 ̄(-7)之间免疫沉淀。SDS—PAGE分析,2株与132K多肽发生特异性反应,只具有HSV-1型特异性,其余3株与120~128K多肽,2株与60K多肽发生特异性反应,具有HSV型共同抗原特异性。7株McAb与CMV均不发生特异反应。这些杂交瘤细胞系的建立对于HSV的临床早期、快速诊断有重要的应用价值。  相似文献   

5.
抗HSV—1杂交瘤细胞系的建立及单抗特性研究   总被引:1,自引:1,他引:0  
以HSV-1为抗原,免疫BALB/C鼠,取其脾细胞在PGE介导下与SP2/0细胞常规融合,IFA法筛选鉴定,反复克隆,共获得7株能稳定、持续分泌抗HSV的McAb的杂交瘤细胞系。核型分析染色体数为87 ̄103条之间,Ig类型分别为IgG1、IgG2b、IgG3,腹水抗体效价在10^-5 ̄10^-7之间免疫沉淀。SDS-PAGE分析,2株与132K多肽发生特异性反应,只具有HSV-1型特异性,其余3  相似文献   

6.
抗胃癌细胞单克隆抗体的制备   总被引:4,自引:0,他引:4  
作者用新鲜胃癌组织匀浆免疫BALB/c小鼠,制备抗胃癌细胞单克隆抗体,一次融合率为92.8%,获得分泌特异性McAb杂交瘤细胞15株,阳性率13.2%,三次克隆纯化后阳性率100%,在体外培养150余天,分泌能力稳定,选择QY-1,QY-2,QY-3,QY-44株McAb进行鉴定,4株McAb对正常纤维组织、红细胞等呈阴性反应,而对胚胎的消化道上皮均呈阳性反应,QY-1 ̄4McAb的胃癌反应阳性率  相似文献   

7.
应用杂交瘤技术,用重组人G-CSF(rhG-CSF)免疫Balb/c小鼠,然后将其脾脏细胞与小鼠骨髓瘤细胞NS-1融合,成功地获得了多种能够分泌抗rhG-CSF单克隆抗体(McAb)的杂交癌细胞。对其中的2株杂交瘤细胞2A6和2C2分泌McAb的研究表明,它们能够特异性地和rhG-CSF结合,而与重组的人GM-CSF、IL-CSF、IL-2、IL-2α和TNF细胞因子无交叉反应,为抗G-CSF特异性McAb。这些杂交瘤细胞经反复冻存和复苏后,仍能稳定地分泌McAb。  相似文献   

8.
作者用新鲜胃癌组织匀浆免疫BALB/c小鼠,制备抗胃癌细胞单克隆抗体(McAb)。一次融合率为92.8%,获得分泌特异性McAb杂交瘤细胞15株,阳性率13.2%,三次克隆纯化后阳性率100%,在体外培养150余天,分泌能力稳定。选择QY-1,QY-2,QY-3,QY-44株McAb进行鉴定。4株McAb对正常纤维组织、红细胞等呈阴性反应,而对胚胎的消化道上皮均呈强阳性反应,QY-1~4McAb的胃癌反应阳性率为94.6~100%,肝癌33.3~60.0%,大肠癌为5.0~35.0%,萎缩性胃炎不伴肠化型为0%,伴肠化型为0~15.0%。本文结果表明QY-1~4McAb相应抗原为非CEA、AFP成份的消化道肿瘤抗原,QY-1~4McAb具有较高的肿瘤特异性。  相似文献   

9.
将用CPV免疫的BALB/c小鼠脾细胞与SPV2/0细胞在聚乙二醇作用下融合,用HI法筛选,有限稀释法克隆三次,得到6株分泌抗CPVMcAb的杂交瘤细胞株,其中4株分泌IgG1,2株分泌IgM。将杂交瘤细胞注射BALB/c小鼠诱生腹水,摸索出了大量生产腹水的最佳实验条件。采用CPV McAb研制触CPV快速诊断试剂盒,检测47份病犬粪样,在30min即可肉眼判定结果,与HA法和常规ELISA相比,具有简便、快速、特异之优点。应用CPVMcAb治疗CPV发病犬287只,治愈率高达97.21%,显著高于其他疗法的治疗效果,治疗机理除对体内病毒的中和作用外,有可能促进机体的主动免疫反应。在对46只幼犬进行CPV疫苗免疫时,使用CPVMcAb的23只,其免疫成功率为100%,而未使用CPVMcAb的23只对照犬,其免疫成功率仅为78.3%,表明CPV McAb在疫苗免疫中可起积极作用。  相似文献   

10.
采用人乳腺癌血清抗原免疫的BALB/c小鼠脾细胞与小鼠骨髓瘤细胞SP2/0融合,经ELISA筛选,有限稀释及克隆后获得了一株稳定分泌特异抗体的杂交瘤细胞株,命名为单克隆抗体GB2(McAbGB2)。McAbGB2经硫酸铵及DEAE纤维素纯化后效价达1∶2×106,经琼脂糖免疫双扩证明McAbGB2属小鼠IgG1亚类。免疫印迹结果表明McAbGB2识别的抗原呈两条区带,分子量分别为116kd及45kd,分布于乳腺癌组织及血清中。  相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

14.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

15.
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.  相似文献   

16.
Objective:To investigate the gene expression of osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) in the bone tissue of patients with hip fracture due to osteoporosis. Methods:OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers(over 50 years old) with hip fracture(Observer Group) and 30 cases of hip facture sufferers with no osteoporosis(Control group) were analyzed with the Semi-Quantitative RT-PCR method. Results:The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion:Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.  相似文献   

17.
Objective:To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas(SPTP). Methods:Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin(HE) staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results:Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin(9/9 cases), AAT(9/9 cases), NSE(9/9 cases), ACT(7/9 cases), CK20(2/9 cases), CgA(1/9 cases), S-100(3/gcases), PR(4/gcases), Syn(3/9 cases) and CD56(5/9cases), negative in CEA and ER. Conclusion:SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.  相似文献   

18.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

19.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

20.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

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