步行量和体成分的中日比较研究

目的 为了解中日儿童及成人的步行和体成分及其相互关系.方法 以中国上海市和日本琦玉县的小学生和成人为对象,采用西铁城 TW02-001电子式步数计测定步行量,FUTREX-5000A体成分测定仪测量体脂百分比.结果 1.中国男女小学生的平均每日步行量分别为12426步和10387步,低于日本小学生(男15 340步,女11 617步),体脂百分比中国男女生分别为21.2%和15.8%,高于日本(男18.9%,女13.1%).中国男女成人的平均每日步行量分别为9826步和8748步,高于日本成人(男:8411步,女7599步);体脂百分比分别为17.5%(男)和25.8%(女),而日本男性为19.8%,女性为26.0%.2.不同活动水平的小学生和成人,其血压和体成分均有差异,步行量高者的血压和体脂百分比都较低.本研究还计算出以肱三头肌皮脂厚度以(X)推测儿童体脂百分比(Y)的回归方程为:Y=10.6646+0.5856X.结论 作为日常生活的步行可促进肥胖发生率的降低.     

薄层扫描法测定食物中胆固醇的含量

本文探讨食物中胆固醇的薄层光密度法的测定。用硅胶Ｇ与０．３％ＣＭＣ－Ｎａ按１：２．２比例制成薄层板，石油醚－乙醚－－冰醋酸（７：３：０．１）为展开剂，５％磷钼酸乙醇溶液显色，日本岛津ＣＳ－９１０型薄层扫描仪定量扫描，测得精神异系数（ＲＤＳ％）为２．９２。稳定性变异系数（ＲＳＤ％）为２．３５，加嘏率在８７．９５－９４．２９％范围内。与气相色谱法（其回收率在９２－１０３％范围）比较，结果相接近，该法简     

粉尘螨主要变应原Derf1基因的改造与可溶性表达

目的在大肠杆菌中提高粉尘螨1类变应原（Derf1）的可溶性表达。方法用RT-PCR方法扩增得到Derf1的全长序列与成熟肽序列（mDerf1）；以已知DerP5基因的前导序列替换Derf1前导序列和原酶序列，重新构建出rDerf1基因；将上述3个基因分别克隆入原核表达载体pGEX-4T-1中表达，经Western-blot对三者的表达产物进行分析鉴定。结果Western-blot表明，pGEX-Derf1与pGEX-mDerf1的表达产物分别为分子量约63000与51000的重组蛋白质，重组蛋白质主要存在于细胞裂解液的沉淀中。pGEX-rDerf1大量表达了可溶性目的蛋白质，分子量约53000，并被成功地分离纯化。以上三种表达产物均可被鼠抗GST抗体与螨过敏患者血清特异地识别。结论表达了含Derf1，mDerf1与rDerf1的三种GST融合蛋白质，它们均具免疫反应性，纯化获得了GST-rDerf1融合蛋白质，为后期的诊断及治疗奠定了基础。     

腹主动脉与其主支夹角的空间定位

甲醛固定的成人尸体32具(男19,女13),造影拍片和实地解剖对照观察,测量腹主动脉主支的发出高度、起点方位、与主动脉的夹角,以及与矢状面的交角。综合以上四项指标,即能构想腹主动脉与其主支夹角的空间位置和变动范围。     

Development of a Western Blot Assay for Detection of Antibodies against Coronavirus Causing Severe Acute Respiratory Syndrome

To identify a major antigenic determinant for use in the development of a rapid serological diagnostic test for severe acute respiratory syndrome (SARS) coronavirus infection and to study the immune response during SARS coronavirus infection in humans, we cloned the full length and six truncated fragments of the nucleocapsid gene, expressed them, and purified them as glutathione S-transferase-tagged recombinant proteins. The reactivities of the recombinant proteins to a panel of antibodies containing 33 SARS coronavirus-positive sera and 66 negative sera and to antibodies against other animal coronaviruses were screened. A truncated 195-amino-acid fragment from the C terminus of the nucleocapsid protein (N195) was identified that had a strong ability to detect antibodies against SARS coronavirus. No cross-reaction was found between the N195 protein and antibodies against chicken, pig, and canine coronaviruses. The N195 protein was used to develop a Western blot assay to detect antibodies against SARS coronavirus in 274 clinically blinded samples. The specificity and sensitivity of this test were 98.3 and 90.9%, respectively. The correlation between our Western blotting assay and an immunofluorescence assay (IFA) was also analyzed. The results of our Western blot assay and IFA for the detection of SARS coronavirus-positive sera were the same. Thus, the N195 protein was identified as a suitable protein to be used as an antigen in Western blot and other possible assays for the detection of SARS coronavirus infection.     

Novel immunofluorescence assay using recombinant nucleocapsid-spike fusion protein as antigen to detect antibodies against severe acute respiratory syndrome coronavirus

Severe acute respiratory syndrome (SARS) is caused by a novel and highly infectious virus named SARS coronavirus (SARS-CoV). Among the serological tests currently available for the detection of SARS-CoV, a whole-virus-based immunofluorescence assay (IFA) was considered one of the most sensitive assays and served as a "gold standard" during the SARS epidemic in Singapore in 2003. However, the need to manipulate live SARS-CoV in the traditional IFA limits its wide application due to the requirement for a biosafety level 3 laboratory and the risk of laboratory infection. Previously, we have identified two immunodominant epitopes, named N195 and Sc, in the two major structural proteins, the N and S proteins, of SARS-CoV (Q. He, K. H. Chong, H. H. Chng, B. Leung, A. E. Ling, T. Wei, S. W. Chan, E. E. Ooi, and J. Kwang, Clin. Diagn. Lab. Immunol., 11:417-422, 2004; L. Lu, I. Manopo, B. P. Leung, H. H. Chng, A. E. Ling, L. L. Chee, E. E. Ooi, S. W. Chan, and J. Kwang, J. Clin. Microbiol. 42:1570-1576, 2004). In the present study, the N195-Sc fusion protein was highly expressed in insect (Sf9) cells infected with a recombinant baculovirus bearing the hybrid gene under the control of a polyhedrin promoter. An IFA based on Sf9 cells producing the fusion protein was standardized with 23 serum samples from patients with SARS, 20 serum samples from patients with autoimmune diseases, and 43 serum samples from healthy blood donors. The detection rates were comparable to those obtained with a commercial SARS-CoV IFA kit (EUROIMMUN, Gross Groenau, Germany) and a conventional IFA performed at the Singapore General Hospital. Our data showed that the newly developed IFA could detect SARS-CoV in 22 of the 23 SARS-CoV-positive serum samples and gave no false-positive results when the sera from patients with autoimmune diseases and healthy individuals were tested. The detection rate was identical to those of the two whole-virus-based IFAs. Thus, the novel N-S fusion antigen-based IFA could be an attractive alternative to present whole-virus-based IFAs for the diagnosis of SARS-CoV infection.     

高频电刀对家兔活体肌肉组织的作用

目的 :探讨高频电刀对肌肉组织结构的作用。方法 :用高频电极在不同输出功率时测定大白家兔活体肌肉组织的电流强度 ,灼伤中心区汽化深度和汽化表面积 ,光镜和电镜下观察创面下 0～ 6mm深度的组织结构变化。结果 :随着电流输出功率的增强 ,肌肉组织的电流强度 ,中心区汽化深度和汽化表面积也随之增加 ,创面深部组织显微和超微结构发生了热凝固性改变 ,损伤程度随着离中心区距离的延长而减弱 ,但在 6mm深处仍有较明显的损伤性改变。结论 :用高频电刀操作时应注意对深部肌肉组织的损伤性作用 ,为减少损伤应注意操作时的电流强度和电灼时间。     

急性鸭乙型肝炎病毒感染病毒清除机理研究

目的 :进一步阐明嗜肝病毒自然感染过程中病毒清除机理。方法 :7只 2～ 3月龄成年重庆麻鸭静脉接种 10～ 2 0mlDHBV阳性血清 (5× 10 8～ 1× 10 9genome) ,接种后每周采血检测外周血中DHBVDNA、DHBsAg和特异抗体的产生 ;感染后第10、35天分离外周血单个核细胞用于抗原特异细胞增殖实验 ;第 5、30、6 0天取肝组织标本进行DHBVDNASouthern杂交、DHB sAg免疫组化及肝组织病理检测。结果 :DHBV感染成年鸭在 1～ 2w潜伏期后出现急性、一过性感染 ,感染高峰期肝内存在多拷贝的DHBV所有复制中间体形式 ,包括cccDNA。进一步分析显示病毒血症的消失是在快速抗原特异细胞增殖反应和高滴度特异抗体产生之后 ;与此同时 ,整个急性感染期间 ,肝细胞并无明显的损害。结论 :非细胞直接损伤机制在嗜肝病毒清除过程中发挥了重要作用。