人CGT52TGT MBL突变体在CHO细胞中的表达及其产物分析

目的: 初步探索MBL基因CGT52TGT点突变引起调理吞噬缺损的机制。方法: 采用PCR技术, 从质粒pMBLm52中获取含CGT52TGT点突变的MBL基因, 将其插入真核表达载体pcDNA4 /HisMaxC中构建重组表达载体。经测序验证后, 电转染入CHO细胞。以800mg/LZeocin筛选转染后的CHO细胞30d; 随后的30d中, 维持Zeocin的浓度在200mg/L, 以获取稳定转染的细胞。以RT PCR分析其mRNA的表达情况。表达产物经Ni NTAagarose纯化后, 以非还原SDS PAGE和Westernblot对表达产物进行初步鉴定。结果:以PCR扩增的MBLm52基因片段长约750bp, 将其插入表达载体构建重组真核表达载体pcDNA4 /HisMaxC MBLm52, 测序验证序列正确后将其电转染入CHO细胞。从细胞培养上清中获得的纯化的表达产物, 主要为相对分子质量(Mr)约60 000的分子, 寡聚化程度明显低于重组人野生型MBL和从人血浆中分离的MBL。结论: MBL基因CGT52TGT点突变可能并不影响其表达产物向胞外分泌的过程, 但突变后产生的Cys可能形成新的二硫键, 影响MBL结构单位和/或寡聚分子的形成, 推测该突变MBL蛋白不能发挥正常的功能。     

Measurement of psychological factors associated with genetic testing for hereditary breast,ovarian and colon cancers

Despite numerous individual studies of psychological factors (depression, anxiety, distress) related to genetic testing for inherited cancer syndromes (CGT), there has been no systematic review of the psychological factors are measured among individuals at increased risk for hereditary breast, ovarian, or colon cancer. Our review provides an analysis of psychological factors in studies of CGT and discusses the instruments most commonly used to measure them. We performed a literature search using three major OVID databases from 1993 to January 2003. In the 19 studies that met our inclusion criteria, the most commonly assessed psychological factors were distress, anxiety, and depression. These factors were most often measured by the impact of event scale (IES), the state-trait anxiety inventory (STAI), and the Centers for Epidemiologic Studies and Depression scale (CES-D), respectively. Our results show deficits in the existing body of literature on psychological factors associated with CGT including limited documentation of psychometrics and variability in instrumentation.     

Calculation of Pesticide Degradation in Decaying Cotton Gin Trash

Pesticide residues were measured in stockpiled cotton gin trash (CGT) over a 2-year period. Samples were analysed by GC/MS/MS and interpretation of the results was aided by the presence of DDE residues, remnant from prior DDT use. Fourteen pesticide residues from current agricultural practice were detected in CGT. Several of these, including indoxacarb, profenofos, chlorpyrifos, propargite, bifenthrin, ethion and cyhalothrin, were more persistent than expected on the basis of published data for soil dissipation. The results showed a complex pattern of pesticide residue decay over time because of the simultaneous decomposition of the CGT matrix.     

Neurocognitive deficits associated with shoplifting in young adults

Objectives

Shoplifting is a relatively common behavior in young adults, but the demographic and neuropsychological correlates of shoplifting remain poorly characterized in this context.

Method

Non–treatment-seeking young adults (18-29 years) were recruited from the general community on the basis of having no Axis I disorders, no history of illicit substance use, and no history of conduct disorder or antisocial personality disorder. Participants were grouped according to presence or absence of shoplifting (at least 1 time over the past 12 months). Measures relating to impulsivity along with objective computerized neuropsychological measures were collected.

Results

Shoplifters (n = 14) and controls (n = 95) did not differ significantly in terms of salient demographic characteristics. Compared with controls, shoplifters endorsed higher impulsivity on the Barratt Impulsiveness Scale and Eysenck Impulsivity Questionnaire, gambled significantly more points on the Cambridge Gambling Task, and showed deficits on the hardest level of difficulty on the Spatial Working Memory task. Performance on executive planning, set-shifting, and response inhibition did not differ significantly between shoplifters and controls.

Conclusions

This study identified significant cognitive deficits in those with past-year shoplifting behavior even in the absence of Axis I disorders and a history of illicit drugs or alcohol. These preliminary findings inform our understanding of the neurocognitive sequelae of shoplifting and its relationship with other impulse control problems, subclinical and clinical. Future work should use longitudinal designs to examine the temporal relationship between these deficits, shoplifting behavior, other impulsive behavior, and functional impairment.     

International Pediatric Otolaryngology Group (IPOG) consensus recommendations: Hearing loss in the pediatric patient

ObjectiveTo provide recommendations for the workup of hearing loss in the pediatric patient.MethodsExpert opinion by the members of the International Pediatric Otolaryngology Group.ResultsConsensus recommendations include initial screening and diagnosis as well as the workup of sensorineural, conductive and mixed hearing loss in children. The consensus statement discusses the role of genetic testing and imaging and provides algorithms to guide the workup of children with hearing loss.ConclusionThe workup of children with hearing loss can be guided by the recommendations provided herein.     

CGT52TGT和GGA57GAA甘露聚糖结合凝集素突变体的构建

目的 构建CGT52TGT和GGA57GAA2种甘露聚糖结合凝集素(MBL)基因突变体。方法 设计2对引物52F／52R和57F／57R(R中引入了点突变)，以含汉族人野生型MBL cDNA的重组质粒pGEM-mbl为模板，采用TaKaRa MutanBEST突变试剂盒进行定点突变，以PER和测序分析进行鉴定。结果 分别用52F／52R和57F／57R为引物对进行PCR，均得到1个约3800bp的DNA片段，自身连接后获得重组质粒。以SP6／T7P为引物对进行PCR，获得约900bp的扩增产物。序列分析表明，2种克隆分别除其第52、57位密码变为TGT、GAA外，其余与野生型MBL cDNA完全相同。结论 构建成功CGT52TGT和GGA57GAA MBL基因突变体，为深入探索MBL基因突变引起调理吞噬缺损的机理和MBL分子的结构-功能关系提供了分子模型。     

益肾通治疗老年慢性肾小球肾炎的临床观察

目的：观察益肾通治疗老年慢性肾小球肾炎和抗衰老的作用机制。方法：口服益肾通煎剂治疗31例老年慢性肾小球肾炎患8周,观察患症状、体征的变化,并测定患治疗前后24h尿蛋白定量、血液流变学及肾功能。结果：益肾通具有改善各项衰老症状,对患24h尿蛋白量、血液流变学及肾功能的影响,与治疗前比较有显性差异（P＜0．05,P＜0．01）。结论：益肾通过老年慢性肾小球肾炎具有较好的综合治疗作用。     

化橘红药材质量评价、监测与应用研究

目的：评价化橘红质量,确定质量监测方法。方法：以形态分类法和显微鉴定法比较原植物及显微构造,以分光光度法比较总黄酮含量,以高效液相色谱法比较柚皮苷含量及指纹图谱,以薄层扫描法比较野漆树苷含量。结果：两类化橘红原植物存在外果皮绒毛、茎非腺毛等显著差异;化橘红总黄酮含量、柚皮苷含量均高于光橘红,野漆树苷含量为光橘红的10倍,指纹图谱也有显著差异。化州柚鲜果重量增加峰值出现在成果41d时,此时其药材质量也较优。结论：道地药材化橘红质量显著优于光橘红,化橘红采收期宜定在41d左右。