弓形虫有效抗原表位的筛选及其对小鼠免疫保护性的研究

目的:利用噬菌体随机肽库技术筛选弓形虫抗原的有效表位，并探讨其免疫保护效果。方法：用纯化的弓形虫免疫兔血清IgC对噬菌体12肽库进行三轮筛选，对获得的日的噬菌体用间接ELISA和Dot—ELISA检测其特异性，并对其中的某些克隆进行Western—blot分析和序列测定；用混和噬菌体克隆及强阳性表位克隆免疫BABL／c小鼠．间接ELISA法测定其特异性抗体滴度，攻击感染后观察小鼠存活情况。结果：经三轮筛选，特异性噬菌体得到富集，随机挑取18个克隆经间接ELISA和Dot—ELISA鉴定，有16个克隆能与弓形虫免疫兔血清IgG呈特异性反应。Western—blot分析显示P2克隆能被免抗弓形虫血清所识别，具有类似于弓形虫抗原的免疫原性，其序列为5’-CTTCAGTTGGATCGGGCTCGGTTTTGGAAT CAGGGT-3’，与GenBank的弓形虫已知序列无一级结构的同源性；与原肽库对照组相比，P2实验组和P总实验组免疫小鼠均能诱导出较高滴度的IgG抗体，抗攻击感染后小鼠的存活率及存活时间也均高于对照组。结论：利用噬菌体随机肽库技术获得了弓形虫抗原的有效模拟睥位，这些表位能诱导对弓形虫的部分保护作用。     

用抗HCV多抗从随机12肽库中筛选抗原表位

目的：用丙型肝炎患者血清中抗丙型肝炎病毒（HCV）抗体从噬菌体随机12肽库中筛选HCV抗原表位。方法：将丙型肝炎患者血清混合,提取纯化的IgG作为固相配基筛选噬菌体随机12肽库,按吸附-洗脱-扩增的淘洗过程进行3轮筛选,随机挑取噬菌体克隆用ELISA法检测其特异性,对4个克隆进行测序。并用ELISA法检测噬菌体克隆的诊断价值。结果：3轮筛选的投入产出比逐轮升高,回收率从(4.6×10-4)%增加到(5.3×10-2)%,具有良好的富集效果。对从第3轮洗脱物中挑选出的18个克隆进行结合试验,发现所挑的克隆与丙型肝炎患者的多克隆抗体有较强的结合力。其中4个克隆测序显示为同一克隆（命名为C1）,其外源插入肽为GSMSPYVRWYTP。用C1检测20例丙型肝炎患者血清的检出率为85.0%。结论：成功地用噬菌体12肽库对丙型肝炎患者血清进行了模拟肽筛选,且得到的模拟肽分子C1具有一定的诊断价值。     

日本血吸虫成虫67kD抗原模拟表位的筛选及其免疫原性

目的：筛选日本血吸虫成虫67kD抗原的模拟表位，对其免疫学活性予以鉴定。方法：用粗提日本血吸虫成虫67kD抗原的抗体IgG作配体筛选噬菌体12肽库，随机挑取三轮筛选后的噬菌体克隆，Dot-ELISA检测其特异性：用混和阳性噬菌体克隆免疫小鼠3次，攻击感染后45d剖杀，计数虫荷。结果：经三轮筛选，特异噬菌体得到富集，挑取的11个噬菌体克隆经Dot-ELISA鉴定均与67kD抗原免疫血清呈阳性反应。混合噬菌体克隆的免疫血清可识别67kD抗原，并具有一定的抗血吸虫的免疫保护效果。结论：利用噬菌体随机肽库技术获得了日本血吸虫成虫67kD抗原的模拟表位，这些表位具有良好的免疫原性。     

噬菌体随机肽库筛选日本血吸虫22.6kDa蛋白有效抗原表位的初步研究

目的：从噬菌体随机12肽库中筛选出中国大陆株日本血吸虫22.6kDa蛋白（Sj22.6kDa抗原表达的短肽分子，探讨其抗血吸虫的交叉免疫保护效果。方法：以纯化的抗日本血吸虫22.6kDa的多克隆抗体IgG为配基，对噬菌体12肽库进行5轮亲和筛选，富集特异性噬菌体，挑取克隆，经序列分析，免疫识别和动物初筛后获得阳性克隆。结果：经过5轮筛选，特异性噬菌体得到有效富集。随机挑选的24个噬菌体克隆中获得4个有效抗原表位。结论：利用噬菌体随机肽库筛选可获得类似日本血吸虫22.6kDa有效抗原表位的短肽分子，这些短肽分子能诱导抗血肿虫的保护性免疫。     

丙型肝炎病毒包膜蛋白E2抗原模拟表位的筛选和鉴定

目的：筛选丙肝病毒（HCV）包膜蛋白（E2）特异性噬菌体模拟表位。方法：以抗－HCV E2的单克隆抗体作为固相筛选分子，对噬菌体同七肽库进行5轮“吸附－洗脱－扩增“的筛选过程，随机挑取50个克隆，经酶联免疫吸附法（ELISA）鉴定、交叉反应以及竞争抑制性实验，最后对所选克隆进行DNA序列分析，以确定HCV E2抗原的模拟表位。结果：经噬菌体富集后，得到12个阳性克隆，确定氨基酸序列XXPXYXW为HCV E2的模拟表位。结论：用噬菌体七肽库成功筛选得到HCV E2的模拟表位，为开展用HCV模拟表位探索HCV的防治研究创造了条件。     

抗大肠杆菌LPS多克隆抗体的制备与LPS模拟肽的筛选

目的获得抗大肠杆菌脂多糖(LPS)多克隆抗体与模拟LPS表位的线性噬菌体展示肽克隆。方法制备兔抗鼠大肠杆菌抗血清,鉴定抗血清效价。以亲和纯化的多克隆抗体为靶,筛选噬菌体随机线性十二肽库。双夹心ELISA和竞争抑制ELISA鉴定阳性噬菌体克隆。结果兔抗血清能与大肠杆菌LPS反应。用亲和层析纯化的多克隆抗体为靶分子进行三轮筛选,随机挑选17个克隆,其中6个克隆显示与多抗结合。大肠杆菌LPS可抑制阳性噬菌体克隆与多抗结合,所有克隆的IC50(达到50%抑制率的LPS浓度)为250ng/ml。结论获得能与大肠杆菌LPS反应兔多克隆抗体,筛选得到可模拟大肠杆菌LPS表位的噬菌体展示线性十二肽。     

从随机多肽库中筛选日本血吸虫抗原模拟表位诱导保护性免疫的研究

目的 从噬菌体随机肽库中筛选出能模拟日本血吸虫抗原表位的短肽分子，并测定其作为疫苗候选分子的潜能。方法 分别用正常及感染日本血吸虫的东方田鼠血清筛选在丝状噬菌体表面表达的随机十二肽库。经三轮亲和筛选后，随机挑取噬菌体克隆用ELISA检测其与日本血吸虫抗血清的反应。用混合噬菌体免疫小鼠并进行攻击感染。结果 随机挑取的12个克隆中，感染东方田鼠血清来源的噬菌体有10个克隆为阳性，正常血清来源的噬菌体有7个克隆为阳性。用感染东方田鼠血清筛到的噬菌体免疫小鼠诱导了部分保护作用（22．6％）及较高的肝卵减少率（68．9％）。然而，正常东方田鼠血清筛到的噬菌体没有保护效果。结论 感染及正常东方田鼠血清筛到的模拟表位具有免疫原性，提示随机肽库技术在日本血吸虫疫苗研制中有其潜在价值。     

应用噬菌体7肽库筛选泡球蚴Em18抗原模拟表位的初步研究

目的：从噬菌体7肽库中筛选泡球蚴Em18抗原模拟表位,为研究和开发新的包虫病诊断抗原提供实验依据.方法：用纯化后的rEm18-GST免疫新西兰白兔,获得抗rEm18-GST的多克隆抗体,进一步纯化,获得抗Em18多克隆抗体IgG,以之作为靶分子,免疫筛选噬菌体随机7肽库.经过5轮的淘筛过程,随机挑取9个蓝色噬菌斑扩增,核苷酸序列测定分析并与Em18进行同源性比较.结果：经5轮筛选后,阳性克隆得到富集,9个噬菌体克隆的氨基酸序列与Em18无同源性.结论：所得肽段可能是Em18抗原模拟表位.     

噬菌体7肽文库筛查类风湿关节炎相关抗体及分析

目的筛选和鉴定与类风湿关节炎(rheumatoid arthritis,RA)患者血清特异性结合的噬菌体7肽,并分析其实际意义。方法采用30例正常人混合血清和30例RA患者混合血清作为筛选配基对噬菌体随机7肽库进行亲和筛选、扩增,获得RA血清特异性结合的噬菌体克隆。用患者混合血清进行Dot-ELISA实验鉴定获得的噬菌体克隆,进一步用RA患者及正常人血清各12例筛选阳性噬菌体的混合克隆。对鉴定的噬菌体克隆进行测序,并分析其同源性。结果获得17个与RA患者混合血清特异性结合的阳性克隆。阳性噬菌体混合克隆与RA患者个体血清反应阳性率明显高于其与正常人血清的反应率。序列分析显示阳性噬菌体克隆的抗原表位之间无共有序列,但与杆菌、球菌等有一定的同源性。结论 RA患者血清中存在与病原体抗原表位结合的抗体成分,提示RA的发病可能与病原体感染有关。     

日本血吸虫尾蚴抗原模拟表位的筛选及其免疫保护性

目的 筛选日本血吸虫尾蚴抗原的模拟表位，探讨其对日本血吸虫的免疫保护性。方法 用粗提日本血吸虫尾蚴抗原的抗体IgC作配体筛选噬菌体12肽库，按“吸附－洗脱－扩增”的过程进行三轮筛选；随机挑取单个噬菌体克隆进行Dot-ELISA检测；用混和噬菌体克隆免疫小鼠3次，攻击感染45d后剖杀冲虫，计算虫数及肝卵数。结果 经三轮筛选，特异性噬菌体得到富集，挑取11个噬菌体克隆经Dot-ELISA鉴定，均与日本血吸虫尾蚴抗原免疫血清呈特异性反应。与对照组相比，混和噬体克隆免疫小鼠的减虫率为18.79%，减卵率为38.00%。结论 利用噬菌体随机肽库技术获得了日本血吸虫尾蚴抗原的模拟表位，这些表位能诱导对日本血吸虫的保护性免疫。     

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.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.     

Possible Spread of adenovirus type 3 from poultry to humans ：indirect evidence from an outbreak in China

Objective:To explore the epidemiology and etiology for an outbreak of acute respiratory tract infection that occurred in one county of Jiangsu Province, China 2004. Methods: Only cases meeting the case definition were included in the study. We reviewed the medical records of the cases who were admitted to the local hospitals, interviewed cases by a standard questionnaire, and then described the epidemiotogic features and analyzed risk factors by means of a case-control study. We collected pharyngeal swab specimens and sent them to different laboratories for isolation and culture. The laboratory used different detection methods such as DIP, PCR, electron microscope examination and microneutralization assay, to identify and then type the positive specimens. Results:A total of 871 cases were reported during the period from April 18 to July 4,2004. The distribution of onset times presented two peaks, one in late May and another in middle June. The epidemic occurred mainly in the elementary and junior high schools in ten townships of one county, and the mean age of the cases was 12 years (range 7 months to 18 years). The course of the disease was acute, and was characterized by fever accompanied with sore throat and tonsillitis. The WBC count of cases was normal or elevated. The mean duration of illness was 5 days (range 2 to 12 days). No fatalities from illness were reported. A case-control study indicated that the possible risk factors were close contact with a case and/or poultry before onset and sharing of towels among members of the family. The typical CPE was observed through inoculating pharyngeal swab specimens into the HEP-2 cell cultures in different laboratories. An infection of adenovirus type 3 was verified by detecting positive specimens in different methods. Conclusion:This investigation demonstrated that the acute respiratory infection in cases was caused by adenovirus type 3. Cases occurred in over 70 schools in ten townships in 2004, and the route of transmission was possibly close contact with cases or droplet transmission.