不同富集培养方法对噬菌体PEf771的滴度影响

目的 对比3种不同富集培养方法对噬菌体PEf771的滴度影响,探讨有效提高噬菌体PEf771滴度的方法。方法 通过噬菌体PEf771/粪肠球菌E. faecalis YN771比例法、挖斑法和丝裂霉素C诱导法在E.faecalis YN771不同生长时期加入PEf771原液进行富集,来获得较高的PEf771的滴度。结果 PEf771/E.faecalis YN771=1∶3和挖斑法得出的PEf771的滴度较高,将这2种方法扩大培养后无明显差异（P> 0.05）,且在24 h滴度达108 pfu/mL。丝裂霉素C诱导法未能明显提高PEf771的滴度。通过在E. faecalis YN771生长早期（OD600=0.3）和对数期（OD600=1）加入PEf771进行富集培养,前者培养6 h滴度高达1010 pfu/ml,24 h滴度仍保持1010 pfu/ml,达到了理想滴度。结论 在E. faecalis YN771培养早期（OD600=0.3）加入PEf771以噬菌体/菌=1∶3的方法或挖斑法富集培养可有效提高PEf771的滴度,为研究E.faecalis YN771的噬菌体PE...     

慢性阻塞性肺疾病急性发作期血清CC10水平与肺炎衣原体感染的相关性研究

目的 探讨慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)急性发作期(acute exacerbation chronic obstructive pulmonary disease,AECOPD)患者血清CC10蛋白(clara cell 10-kD protein),含量与肺炎衣原体感染的关系及其临床意义.方法 采用酶联免疫吸附(ELISA)方法检测71例AECOPD患者及30名正常人血清的CC10水平,收集诱导痰作涂片及培养检测细菌及真菌感染,同时应用血清方法检测肺炎衣原体感染血清抗体滴度.结果 与合并细菌及真菌感染的AECOPD患者相比,合并肺炎衣原体感染者血清CC10的下降程度较高(P＜0.05,P＜0.01),血清CC10水平与肺炎衣原体IgG抗体滴度呈负相关(P＜0.05,r=-0.654).结论 AECOPD患者血清CC10蛋白含量的下降程度与肺炎衣原体急性感染强度呈负相关.     

实验感染青海田鼠血清中鼠疫FI抗体的观察

鼠疫菌实验感染青海田鼠28只,从1-30天取血检测鼠疫FI抗体滴度,其中16份抗体阳性,滴度1: 20-1:640。     

重组腺病毒Ad-sTNFRI-IgGFc转染人气道上皮细胞及其表达的实验研究

目的 构建携带人可溶性肿瘤坏死因子受体I胞外区和IgGFc融合基因的重组腺病毒载体(Ad-sTNFRI-IgGFc),体外转染正常人气道上皮细胞株(HBE135-E6E7),探讨重组腺病毒Ad-sTNFRI-IgGFc转染气道上皮治疗支气管哮喘的可行性.方法 将sTNFRI-IgGFc基因插入腺病毒穿梭质粒pDC316,与辅助质粒pBHGlox△E1,3Crc共转染HEK293细胞,重组产生Ad-sTNFRI-IgGFc,PCR鉴定毒种正确后,进行扩增、纯化和滴度测定,转染培养HBE135-E6E7,流式细胞术检测转染效率,RT-PCR、免疫组化检测转染后细胞中sTNFRI-IgGFc的表达和转录.结果 成功构建了Ad-sTNFRI-IgGFc,感染性滴度达3×1010TCID50/ml,转染后HBE135-E6E7在mRNA和蛋白水平均有sTNFRI-IgGFc表达.结论 构建的Ad-sTNFRI-IgGFc可有效转染HBE135-E6E7,并在其中高效表达,并能拈抗TNF活性,为将表达sTNFRI-IgGFc腺病毒基因治疗哮喘的实验研究提供了基础.     

妊娠期甲状腺过氧化物酶抗体滴度与不良妊娠结局的关系

目的 探讨妊娠期妇女体内甲状腺过氧化物酶抗体(TPOAb)滴度与不良妊娠结局的关系.方法 根据TPOAb滴度水平高低不同将孕期TPOAb阳性的104例孕妇(病例组)分为3组,其中6.0～ 500μIU/mL组43例,501～1 000 μIU/mL组35例,＞1 000 μIU/mL组26例,正常对照组100例,分别统计各组不良妊娠结局发生率,并进行相关统计学分析.结果 正常对照组100例不良妊娠结局发生率为21.0％,病例组104例不良妊娠发生率为44.23％,与正常对照组比较,病例组不良妊娠结局发生率显著增加,差异有统计学意义(P＜0.01).TPOAb不同滴度的不良妊娠结局发生率分别为44.19％、42.88％和46.15％,差异无统计学意义(P＞0.05).结论 孕期TPOAb呈阳性的孕妇其不良妊娠结局发生率升高,但TPOAb滴度水平与不良妊娠结局发生率之间无相关性.     

Mycoplasma pneumoniae pneumonia in hospitalized children diagnosed at acute stage by paired sera

Background Mycoplasma pneumoniae （M. pneumoniae） is a frequent cause of respiratory tract infections. However,there is deficient knowledge about the clinical manifestations of M. pneumoniae infection. We described the clinical and laboratory findings of M. pneumoniae pneumonia in hospitalized children who were all diagnosed by a ≥ fourfold increase in antibody titer.Methods M. pneumoniae antibodies were routinely detected in children admitted with acute respiratory infection during a one-year period. The medical history was re-collected from children whose M. pneumoniae antibody titer increased≥fourfold at the bedside by a single person, and their frozen paired serum samples were measured again for the M.pneumoniae antibody titer.Results Of the 635 children whose sera were detected for the M. pneumoniae antibody, paired sera were obtained from 82 and 29.3% （24/82） showed a ≥ fourfold increase in antibody titer. There were 24 cases, nine boys and 15 girls, aged from two to 14 years, whose second serum samples were taken on day 9 at the earliest after symptom onset; the shortest interval was three days. All children presented with a high fever （≥38.5℃） and coughing. Twenty-one had no nasal obstruction or a runny nose, and five had mild headaches which all were associated with the high fever. The disease was comparatively severe if the peak temperature was ＞39.5℃. All were diagnosed as having pneumonia through chest X-rays. Four had bilateral or multilobar involvement and their peak temperatures were all ≤ 39.5℃. None of the children had difficulty in breathing and all showed no signs of wheezing.Conclusions The second serum sample could be taken on day 9 at the earliest after symptom onset meant that paired sera could be used for the clinical diagnosis of M. pneumoniae pneumonia in children at the acute stage. M. pneumoniae is a lower respiratory tract pathogen. Extrapulmonary complications were rare and minor in our study. High peak temperature （＞39.5℃） is correlated with the severity of M. pneumoniae pneumonia in children.     

EB病毒与系统性红斑狼疮的关联研究

目的：通过检测系统性红斑狼疮（SLE）患者体内的EB病毒DNA载量及病毒衣壳相关抗体（包括IgA及EBV-VCA-IgA）,探究SLE患者体内EB病毒感染情况及EB病毒再次感染对SLE活动度的影响。方法选取100例SLE患者、80例非SLE风湿患者及40例正常人血标本,应用荧光定量聚合酶链反应（PCR）测定EBV-DNA的载量,应用酶联免疫吸附试验（ELISA）测定EB病毒相关抗体。比较各组EB病毒抗体抗体滴度。结果 PCR测定EBV-DNA载量：SLE组与非SLE组的EBV-DNA检出率分别为29.0%（29/100）、11.3%（9/80）,差异有统计学意义（P<0.05）。病毒载量异常组的SLE患者的SLE疾病活动度（SLEDAI）为（15±3）,病毒载量正常患者SLEDAI为（13±2）,差异无统计学意义。异常病毒载量患者与正常者的SLEDAI比较,差异无统计学意义。SLE组的病毒相关抗体EBV-IgA与IgG的抗体滴度高于对照组,且差异有统计学意义（P<0.05）。结论 SLE患者体内EBV抗体及DNA表达较高,提示EB病毒与SLE关系密切,但EBV-IgA或DNA载量对SLE疾病活动度无影响。     

某公司国产流行性感冒裂解疫苗安全性和免疫原性研究

[目的]评价某国产流行性感冒裂解疫苗在儿童和成人中应用的安全性及免疫原性,为推广国产流感疫苗的预防接种提供科学依据. [方法]选择6月龄以上健康人群120人作为受试人群,分成6个年龄组,于上臂三角肌肌内接种某公司流行性感冒裂解疫苗,考核和评价疫苗的安全性和免疫原性. [结果]实验疫苗接种首针后全身及局部反应发生率分别为5%及4.17%,该疫苗在18～60岁人群中和≥60岁人群中分型抗体阳转率均≥50%,18～60岁人群和≥60岁人群分型抗体GMT增长＞4.0倍,18～60岁和≥60岁易感者保护率均≥90%.[结论]本次研究结果与诸多同类疫苗临床反应率大致相同,提示该疫苗具有较好的临床安全性.免疫效果达到欧盟流感疫苗临床试验评价标准.     

实验感染青海田鼠血清中鼠疫FI抗体的观察

[目的]观察青海田鼠血清中鼠疫FI抗体动态.[方法]间接血凝试验.[结果]实验感染青海田鼠28只,16份阳性,滴度1：20～1：640.[结论]青海田鼠对鼠疫菌能产生免疫力.     

拱墅区2005年麻疹疫苗效价及免疫成功率调查

随着麻疹疫苗接种率的逐年提高，拱墅区麻疹发病率也逐年下降，但在2005年麻疹发病率骤升至21．65／10万，为了评价拱墅区的冷链系统和麻疹疫苗接种的效果，开展了疫苗滴度与免疫成功率检测，为我区评价冷链系统和免疫预防工作提供依据。