应用多重连接探针扩增技术快速高通量检测染色体非整倍体

目的评价多重连接探针扩增技术(multiplex ligation-dependent probe amplification,MLPA)在染色体非整倍体诊断中的应用价值。方法应用MLPA技术检测了150例羊水标本、50例外周血标本,所有标本均进行常规染色体核型分析,应用Cof-falyser9.0MLPA-DATA数据分析软件获得MLPA结果,比对MLPA和染色体核型分析结果的准确性,评价两种技术的符合率,总结MLPA技术临床应用过程中的关键要点。结果MLPA扩增后探针信号强度与质控相比比率大于1.3判定为重复,小于0.7判定为缺失。150例羊水标本,MLPA显示非整倍体染色体数目异常14例,与羊水染色体培养核型分析结果相同;50例外周血标本,MLPA异常21例,与培养结果符合率100%。DNA质量和浓度是实验成败的关键,Coffalyser9.0MLPA-DATA数据分析软件可以实现MLPA数据处理通量化。结论MLPA技术用于普通的染色体非整倍体数目检测快速、特异、敏感,弥补了常规染色体培养周期长等缺点,对于大量的产前诊断羊水标本,可以实现高通量检测,具较高的临床应用价值。     

尿液分析在健康体检中的重要性

尿液是人体代谢的重要产物,它在体内产生的全过程相当复杂,不仅有泌尿系统的所有器官参与,而且许多重要器官也会协同工作.所以,尿液的变化不但能反映泌尿系统各重要器官的情况,还能看出其他有关器官是否正常.尿液检查是一种无创伤性检查,应用于健康人普查,易取得满意的效果,有助于发现亚健康人群,进行早期诊断及疾病预防.本文对5 292例健康体检者尿液进行了分析,实验结果证明尿液分析项目在健康体检中有非常重要的作用.     

铃铛刺花提取物急性毒性及抗疲劳作用研究

目的：研究铃铛刺花提取物对实验小鼠的急性毒性作用及抗疲劳作用。方法选择SPF级KM种小鼠为受试对象,采用急性毒性试验测定最大耐受法,采用抗疲劳试验评价其抗疲劳的功效,为其进一步应用的安全性提供理论依据。结果最大耐受量为＞120g· kg－1流浸膏,相当于人临床每天口服用量的300倍;与空白对照组比较,铃铛刺花提取物中剂量和高剂量能明显延长小鼠的爬杆时间和小鼠游泳时间,具有统计学意义（P＜0.01）。铃铛刺花提取物中剂量和高剂量能显著降低运动后小鼠血乳酸含量,并且能显著提高运动后小鼠肝糖原的含量,具有统计学意义（P＜0.01）。铃铛刺花提取物可以将降低运动后小鼠尿素氮的含量,但无统计学意义。结论在本次实验研究条件下铃铛刺花提取物的MTD＞120g生药／kg小鼠,实属无毒级,并且铃铛刺花提取物可显著减少运动后乳酸的堆积,提高机体对抗疲劳的能力;可显著提高运动后小鼠肝糖原水平,增加有氧运动糖份供给,从而提高小鼠的抗疲劳作用。     

应用多重连接依赖式探针扩增技术快速高通量诊断胎儿染色体非整倍体异常

目的 评价多重连接依赖式探针扩增技术(multiplex ligation-dependent probe amplification,MLPA)在染色体非整倍体诊断中的应用价值,为我国羊水染色体诊断提供一种快速、特异、高通量的分子诊断手段.方法 应用MLPA技术检测了500份羊水标本,所有标本均进行荧光原位杂交(fluorescence in situ hybridization,FISH)技术检测和常规染色体核型分析,应用RH-MLPA-v511数据分析软件获得MLPA结果,比较MLPA技术与FISH和染色体核型分析结果的准确性,总结MLPA技术临床应用过程中的关键要点.结果 在500份羊水标本中,MLPA检测成功率97%.3个工作日完成结果的为92%,需重复检测的为5%,失败为3%.对染色体非整倍体异常检测敏感性和准确性100%.证实38例非整倍体病例探针信号比值＞正常二倍体4s,2例疑似三体结果＞2s.分析了21号染色体8条探针的杂交效率,21三体患者8条探针中平均4条探针比值＞1.3.结论 MLPA技术具有快速、特异、敏感、高通量、成本低等特点,可用于产前染色体非整倍体数目的快速检测,是传统染色体培养方法的补充,临床应用价值较高.

Abstract：

Objective To assess the diagnostic value of multiplex ligation-dependent probe amplification (MLPA) for detection of common chromosome aneuploidy in amniotic fluid (AF) cells in order to obtain an accurate, rapid, cost-effective and high-throughput method in routine prenatal clinical practice.Methods The MLPA test was performed on 500 AF samples by using kit P095 and the results were obtained by using analysis software RH-MLPA-v511. The results were compared with that from fluorescence in situ hybridization (FISH) and traditional karyotyping (TK). The technical critical issues were analyzed in routine diagnostic application. Results The absolute specificity and sensitivity of the MLPA test to detect the aneuploidy were 100%. For the 500 AF samples, the success rate of the MLPA tests was 97%. Among them 92% were finished within three working days and 5% required more days for repeating. The test failure rate was 3%. The results confirmed that for the 38 detectable aneuploid samples,the probe reliability weighted mean ratio values were more than 4SD compared to normal diploids and the 2 suspected trisomy samples were more than 2SD. In this study, authors analyzed hybridization efficiencies of 8 probes for chromosome 21, and the presence of a trisomy was considered if at least 4 of the 8 probes gave probe ratio of ＞1.3. Conclusion The data suggested that MLPA is a rapid, simple and reliable method for large scale testing for aneuploidy of chromosomes 13, 18, 21, X, or Y in AF. The MLPA technology is complementary to AF culture and valuable for prenatal diagnosis.     

应用多重连接依赖式探针扩增技术快速高通量诊断胎儿染色体非整倍体异常

Objective To assess the diagnostic value of multiplex ligation-dependent probe amplification (MLPA) for detection of common chromosome aneuploidy in amniotic fluid (AF) cells in order to obtain an accurate, rapid, cost-effective and high-throughput method in routine prenatal clinical practice.Methods The MLPA test was performed on 500 AF samples by using kit P095 and the results were obtained by using analysis software RH-MLPA-v511. The results were compared with that from fluorescence in situ hybridization (FISH) and traditional karyotyping (TK). The technical critical issues were analyzed in routine diagnostic application. Results The absolute specificity and sensitivity of the MLPA test to detect the aneuploidy were 100%. For the 500 AF samples, the success rate of the MLPA tests was 97%. Among them 92% were finished within three working days and 5% required more days for repeating. The test failure rate was 3%. The results confirmed that for the 38 detectable aneuploid samples,the probe reliability weighted mean ratio values were more than 4SD compared to normal diploids and the 2 suspected trisomy samples were more than 2SD. In this study, authors analyzed hybridization efficiencies of 8 probes for chromosome 21, and the presence of a trisomy was considered if at least 4 of the 8 probes gave probe ratio of ＞1.3. Conclusion The data suggested that MLPA is a rapid, simple and reliable method for large scale testing for aneuploidy of chromosomes 13, 18, 21, X, or Y in AF. The MLPA technology is complementary to AF culture and valuable for prenatal diagnosis.     

多元化教学方法在天然药物学基础教学中的应用

结合中职药剂专业天然药物学基础课程教学内容特点,根据不同的教学环节总结课堂理论教学、课堂实验教学、户外实践教学方法.     

应用多重连接探针扩增筛查高风险胎儿染色体亚端粒区变异北大核心CSCD

目的探讨多重连接探针扩增（multiplex ligation—dependent probe amplification,MLPA）技术在产前大规模筛查染色体亚端粒区变异中的应用。方法选取1050例高风险胎儿,抽取羊水或脐带血,进行核型分析和MLPA检测。染色体亚端粒区变异结果用染色体微阵列验证。结果核型分析发现染色体非整倍体23例,末端异常8例。MLPA检出了所有核型分析发现的染色体非整倍体和末端异常,并对4例染色体末端异常提供了更清晰的描述。此外,MLPA发现5例核型分析为正常的样本存在染色体亚端粒区变异,经微阵列验证2例为假阳性结果,假阳性率为0．19％。MLPA实际检出染色体亚端粒区变异11例,检出率为1．05％。结论将MLPA技术应用于大规模筛查高风险胎儿的染色体异常,可以缩短检测周期,发现亚微观染色体变异,为遗传咨询和产前诊断提供依据。     

中职药学专业药品营销方向学生核心技能考核方案研究

针对药剂专业药品营销方向药品零售岗位制订核心技能考核方案,规范学生的操作,加强学生顶岗实际操作的能力,凸显工学结合的教育理念。     

中职药剂专业开设综合性实验的探索

在药剂专业的课程设置中,实验教学占了很大比例。针对传统实验教学中的不足,在药剂专业的实验教学中增设综合性实验,有助于培养学生的职业素质和职业能力。     

“工学结合”与中职药剂人才培养模式的探索

药剂中等职业教育可设置的专业岗位群有制剂技术岗位群、药品营销岗位群、医院调剂员岗位群,工学结合人才培养模式突出能力培养,现从行业背景、专业设置、人才培养模式、教学体系等方面入手对中职药剂人才培养模式进行探索。