人心肌肌钙蛋白I单克隆抗体的制备及鉴定

目的 :制备抗人心肌肌钙蛋白 I(c Tn I)单克隆抗体 (c Tn I Mc Ab)并进行初步鉴定。 方法 :从健康意外死亡者的心肌中提取、纯化人 c Tn I,以其为抗原 ,免疫 BAL B/ c小鼠后 ,取其脾淋巴细胞与小鼠 Sp2 / 0骨髓瘤细胞融合 ,反复筛选及克隆 ,得到能稳定分泌 c Tn I Mc Ab的杂交瘤细胞株。初步鉴定 c Tn I Mc Ab。结果 :经间接 EL l SA法筛选 ,3次克隆化后获得 5株能稳定分泌 c Tn I Mc Ab的杂交瘤细胞 (3A7、3A 11、3D2、3F10和 1H9) ,5株单抗免疫球蛋白亚类鉴定均为 Ig G2 a类。染色体数目 92～ 110条。 5株单克隆抗体腹水按 1∶ 10 0稀释 ,与 L DH、CK、CK- MB、AST和人心肌肌钙蛋白 T(c Tn T)行交叉反应为阴性。 5株 c Tn I Mc Ab腹水效价为 3.2× 10 - 6 ～ 1.6× 10 - 7。c Tn I Mc Ab相加试验表明 ,3D2、3F10与 3A7、3A11、1H9能识别不同的抗原表位。 结论 :本实验成功制备了具有较高活性的 c Tn I Mc Ab,具有良好应用开发价值。     

柯萨奇B_3病毒单克隆抗体的制备及鉴定

用柯萨奇 B3病毒免疫 Balb/c小鼠。取脾细胞与 Sp2 /0细胞进行融合及细胞培养。用间接免疫荧光法筛选阳性细胞。并用 ELISA法等行 Ig类及亚类的鉴定。结果 :经融合、筛选、克隆化后共获得 2 6株稳定分泌特异性 Mc Ab的杂交瘤细胞 ,其中 3株分泌Mc Ab为 Ig G1类 ,2株为 Ig G2 b类 ,4株为 Ig G2 a类 ,1 7株为 Ig G3类。 2 6种 Mc Ab中除少数杂交瘤株的 Mc Ab能与同组不同型发生交叉反应及 Mc Ab2 B2 与 Polio 型、Mc Ab1 H10 与CVA4 有交叉反应外 ,其余的 Mc Ab仅与本型的病毒起反应。提示 CVB3- Mc Ab具有高度的特异性及均一性 ,可广泛应用于基础医学及临床医学研究     

Monoclonal anti-idiotype antibody bearing the internal image of nasopharyngeal carcinoma associated antigen

目的　制备和鉴定具有鼻咽癌相关抗原内影像的抗独特型单克隆抗体 (Ab2 )。方法　用鼻咽癌单抗 (Ab1)免疫小鼠 ,免疫鼠脾细胞与骨髓瘤细胞SP2 / 0融合获得杂交瘤细胞系。用SandwichELISA和结合抑制试验筛选阳性克隆。为进一步证实Ab2是否具有鼻咽癌相关抗原的内影像 ,用Ab2免疫小鼠获得抗血清 ,以ELISA和竞争抑制试验检测Ab3。用迟发性变态反应和T细胞增殖试验检测Ab2诱导的细胞免疫反应。结果　选用两株抗鼻咽癌单抗FC2、HNL5 (Ab1)为免疫原 ,制备两株抗相应Ab1独特型的抗独特型抗体 2H4和5D3(Ab2 )。 2H4、5D3能分别与FC2、HNL5特异性结合 ,并能抑制相应Ab1与靶细胞HNE2的反应 ,诱导产生能与相应Ab1竞争结合靶抗原的抗抗独特型抗体 (Ab3)。并能诱发特异性迟发型超敏反应 (DTH)和细胞增殖反应。结论　抗独特型抗体 2H4、5D3具有鼻咽癌相关抗原的内影像 (Ab2 β) ,能模拟鼻咽癌相关抗原诱导体液和细胞免疫反应。     

大规模接种甲肝减毒活疫苗(H2株)后人群的免疫持久性

目的评估甲肝减毒活疫苗(H2株)大规模接种后人群的免疫持久性以达到控制甲肝流行的目的。方法选择220名免前抗甲肝抗体阴性的儿童(年龄1～3岁)作血清学追踪观察,分别于接种H2株疫苗后的2月、12月、6年和10年观察抗甲肝抗体的阳性率和几何平均滴度。另外根据随机原则选择不同年龄组的人群(3岁、6岁、9岁、15岁、18岁、25岁和35岁)比较疫苗接种前和接种后10年抗甲肝抗体的阳性率。10年流行病学追踪观察疫苗接种率和甲肝发病率之间的关系。血清检测方法用ELISA法(用WHO标准品校正)和ABBOTT(美国)AxsymmEIA法。结果在血清学追踪观察人群中疫苗接种后2个月和接种后10年,抗体阳转率分别为98.6%和80.2%;18岁以下年龄组疫苗接种前和疫苗接种后10年抗体阳性率差异有显著性意义,分别从7.69%升高到70.45%(3岁组)、52.58%升高到71.78%(18岁组)。流行病学效果观察:当疫苗接种率达到57%～74%时,2个试验点无甲肝流行;当疫苗接种率达到85%时,2个试验点无甲肝病例的发生;当疫苗接种率保持在85%～91%时,1～15岁年龄组儿童连续9年没有甲肝病例发生。结论通过10年观察,甲肝减毒活疫苗大规模的免疫接种在预防和控制甲肝的流行与发生过程中具有显著的免疫持久性。     

具有鼻咽癌相关抗原内影像的抗独特型抗体研究

用抗人鼻咽癌单抗FC2 ,HNL5 (Ab1)分别免疫Balb/C小鼠 ,制备了两株抗独特型抗体 2H4和 5D3(Ab2 )。结果显示 :2H4和 5D3分别与FC2和HNL5特异性结合 ,抑制Ab1与靶细胞的反应 ,诱导产生能与相应Ab1竞争结合靶抗原的抗抗独特型抗体 (Ab3) ;并能诱发特异性迟发型超敏反应 (DTH)和细胞增殖反应。表明 2H4和 5D3具有鼻咽癌相关抗原的内影像 (Ab2 β) ,能模拟鼻咽癌相关抗原诱导体液和细胞免疫反应。     

半乳糖基抗CD3单抗的制备及体内趋肝性研究

目的　了解半乳糖基抗 CD3 单抗 - TIL 复合物 (Gal- Anti- CD3 - Mc Ab- TIL)的体内趋肝细胞性。方法　制备半乳糖基抗 CD3 单抗 (Gal- Anti- CD3 - Mc Ab) ,采用苯酚 -硫酸法测其糖密度 ;经动物外周静脉分别注入用 1 2 5 I标记的抗 CD3 单抗 (Anti- CD3 - Mc Ab)和用 1 3 1 I标记的 Gal- Anti- CD3 - Mc Ab,分别测定两者在动物体内各脏器的放射活性。结果　 Gal- Anti- CD3 - Mc Ab的糖密度值为 5 8.12 ;外周静脉注射给药 ,Anti- CD3 - Mc Ab趋于肺内浓聚 ,而 Gal- Anti- CD3 - Mc Ab有显著的趋肝性 ,且于靶器官有较长时间停留。结论　经外周静脉途径注射 ,Gal- Anti-CD3 - Mc Ab有显著的体内趋肝细胞性 ,其中半乳糖基与肝结合蛋白所介导的受体 -配体反应在 Gal- Anti- CD3 - Mc Ab的趋肝细胞过程中发挥主导作用     

具有鼻咽癌相关抗原内影像的抗独特型抗体研究

用抗人鼻咽癌单抗FC2,HNL5(Ab1)分别免疫Balb／C小鼠,制备了两株抗独特型抗体2H4和5D3(Ab2)。结果显示：2H4和5D3分别与FC2和HNL5特异性结合,抑制Ab1与靶细胞的反应,诱导产生能与相应Ab1竞争结合靶抗原的抗抗独特型抗体(Ab3);并能诱发特异性迟发型超敏反应(DTH)和细胞增殖反应。表明2H4和5D3具有鼻咽癌相关抗原的内影像(Ab2β),能模拟鼻咽癌相关抗原诱导体液和细胞免疫反应。     

甲型肝炎减毒活疫苗（H2减毒株）接种后的中和抗体测定

采用甲型肝炎减毒活疫苗(H_2减毒株)接种者的双份血清与H_2减毒株(中和用)的病毒,根据ELISA双夹心捕捉原理建立了甲型肝炎中和抗体的测定方法。结果证明接种疫苗后产生的抗HAV抗体为中和抗体,1∶2稀释血清至少可以中和10~3TCID_(50)HAV。对此特异性抗体测定有助于评价甲肝减毒活疫苗的免疫效果。     

卵巢癌抗抗独特型单克隆抗体3D12的制备及初步研究

目的:制备卵巢癌抗抗独特型单克隆抗体并进行初步鉴定,为深入研究卵巢癌疫苗的主动免疫机制奠定基础.方法:以卵巢癌抗独特型单克隆抗体6B11作为免疫原,采用杂交瘤技术制备抗抗独特型单克隆抗体3D12.经酶联免疫吸附实验(ELISA)、免疫组织化学、流式细胞学、免疫印迹杂交等检测其特异性结合抗原的性质;竞争抑制实验鉴定其特异性结合的抗原表位.结果:获得1株稳定分泌3D12的杂交瘤细胞,所分泌的3D12能够特异性结合6B11及卵巢癌初始抗原OC166-9,并能竞争抑制COC166-9(抗OC166-9的单克隆抗体Ab1)与OC166-9的结合.免疫印迹杂交显示,3D12与COC166-9免疫沉淀的蛋白的相对分子质量相同.该抗体为IgM,亲和力约为8.34×10⁷ L/mol.结论:卵巢癌抗抗独特型单克隆抗体3D12属Ab1样Ab3,间接证实6B11为抗原内影像型抗独特型抗体.     

抗人AFP单链抗体基因的构建和在大肠杆菌中的表达

目的　构建抗人 AFP单链抗体 (Sc Fv)的基因并转入大肠杆菌 BL - 2 1(DE3)中诱导表达。分离纯化Sc Fv并鉴定其生物活性。方法　用 RT- PCR方法从能分泌特异性抗人 AFP单克隆抗体的杂交瘤细胞中分离纯化抗体 VH和 VL基因。用重叠延伸 PCR方法将 VH和 VL拼接在一起 ,构建抗人 AFP单链抗体的基因。将 Sc Fv基因克隆至 p GEM- T载体构建 ,用限制性内切酶切以及测序鉴定。将 Sc Fv基因连接到 p ET32 (a )质粒 ,转化 BL - 2 1(DE3)细菌。抗性生长的克隆用异丙基硫代 -β- D-半乳糖苷 (IPTG)诱导 4 h。溶解细菌并纯化 Sc Fv,通过 SDS-PAGE电泳及竞争抑制 EL ISA实验检测其活性。结果　获得的 VH含 339bp,来自小鼠 Ig Gγ链 ,VL含 312 bp,来自小鼠κ链。 VH和 VL通过编码 (G4 S) 3连接肽的 4 5 bp序列连接在一起构成含 6 96 bp的 Sc Fv基因。 BL - 2 1(DE3)中表达的 Sc Fv抗体与融合蛋白 (Trx A)融合在一起并以包涵体的形式存在。 Trx A- Sc Fv相对分子质量约 4 0× 10 3,Sc Fv相对分子质量约 2 4× 10 3。竞争 EL ISA实验显示 :Trx A - Sc Fv可抑制 4 1%的 Mc Ab与抗原结合 ,而Sc Fv抗体则可抑制 5 3%的 Mc Ab与抗原结合。结论　我们成功构建了由 6 96个碱基组成的抗人 AFP单链抗体的基因 ,并在大肠杆菌获得了     

Value of D-Dimers in patients with acute aortic dissection

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.     

Research of combined liver-kidney transplantation model in rats

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

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.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

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.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

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.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

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.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

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.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

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.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.