不同引物标记物在显色法芯片检测结核分支杆菌利福平和异烟肼耐药基因中应用比较

目的比较不同标记物在显色法芯片检测结核分支杆菌利福平和异烟肼耐药基因中的影响。方法分别采用地高辛、荧光素和生物素标记引物,建立地高辛-抗地高辛、荧光素-抗荧光素和生物素-亲和素酶呈色法芯片检测结核分支杆菌利福平(RFP)和异烟肼(INH)耐药基因方法,同时用3种方法检测46株结核分支杆菌临床分离株、19种非结核分支杆菌标准株、9种非分支杆菌标准株和结核分支杆菌H37Rv标准株,并对3种方法的敏感性和重复性进行比较。结果经3种方法平行检测,46例结核分支杆菌临床分离株结果完全一致;19种非结核分支杆菌标准株、9种非分支杆菌标准株结果均为阴性,结核分支杆菌H37Rv标准株结果均为野生型;地高辛系统和荧光素系统敏感度为2.0×103拷贝/ml,生物素-亲和素系统敏感度为2.0×104拷贝/ml;3种方法均有良好的重复性。结论3种方法均具有良好的重复性和特异性,生物素系统敏感度略低于地高辛系统和荧光素系统。     

Exposure to monocrotophos pesticide during sexual development causes the feminization/demasculinization of the reproductive traits and a reduction in the reproductive success of male guppies (Poecilia reticulata)

Monocrotophos is a highly toxic organophosphorus pesticide that has been confirmed to be an endocrine‐disrupting chemical. To evaluate the influence of this pollutant on the reproductive system of male fish, we studied the sex steroid levels, reproductive traits, sex ratio, and reproductive success in male guppies (Poecilia reticulata) exposed to 40% monocrotophos pesticide at the nominal concentrations of 0.01, 0.10, and 1.00 mg/L for 90 days from birth to adulthood in a semi‐static exposure system. Radioimmunoassay and western blot analyses demonstrated that the long‐term exposure to monocrotophos pesticide during the sexual development of male guppies caused a significant increase in 17β‐estradiol levels and consequently induced vitellogenin synthesis, suggesting the feminization of the males. Monocrotophos pesticide also caused a significant decrease in testosterone levels, which consequently inhibited testis growth and reduced the sperm count and the area and intensity of their sexually attractive orange spots, which collectively indicated the significant demasculinization of the male sexual characteristics. Furthermore, these changes in the sexual characteristics at the cellular and organ levels translated into ecologically important effects on the reproductive success at the individual level, as measured by a decrease in offspring production and survival rate. The present study provides the first evidence that monocrotophos pesticide can cause severe reproductive abnormalities in fish due to its endocrine‐disrupting action.     

显色法芯片检测结核分枝杆菌利福平和异烟肼耐药基因

目的建立显色法芯片检测结核分枝杆菌耐药基因的方法。方法设计4对地高辛标记引物,扩增结核分枝杆菌rpoB、katG、inhA和ahpC 4个基因部分片段,根据结核分枝杆菌利福平(RFP)和异烟肼(INH)4条耐药相关基因上8个位点的25种单核苷酸多态性设计探针制作芯片,扩增产物与芯片杂交,显色法判断结果;用该法检测46株结核分枝杆菌临床分离株。结果扩增产物琼脂糖电泳可见4条大小分别为165、181、245、315 bp DNA条带;结核分枝杆菌菌液浓度为1．6×103／ml时,该法仍可检测到各位点野生及突变信号;19种非结核分枝杆菌标准株和9种非分枝杆菌标准株扩增产物无DNA条带,与芯片杂交亦无信号,H37Rv结核分枝杆菌标准株芯片检测各位点均为野生型;5株结核分枝杆菌临床分离株重复检测5次,结果完全一致;6份PCR产物的测序结果与芯片检测结果完全一致;46株结核分枝杆菌临床分离株中,RFP耐药株28株,芯片检出突变株24株,突变率为85．7％,RFP敏感株18株,芯片检出突变株2株。INH耐药株31株,芯片检出突变株20株,突变率为64．5％,INH敏感株15株,芯片检出突变株4株。结论显色法芯片检测结核分枝杆菌耐药基因具有较高的敏感性和特异性,无需特殊仪器设备,有一定的推广应用价值。     

氯己定溶液与含氯消毒剂对医用织物着色作用的探讨

目的 调查医用织物洗涤后着色的原因,寻找避免织物着色的方法.方法 接触氯己定后再与含氯消毒剂接触的医用织物为试验组,接触乙醇、洗衣粉再接触含氯消毒剂为对照组;通过不同的接触与洗涤方法,观察各组着色及褪色情况.结果 试验组接触250～500 mg/L次氯酸钠后1 min即出现着色,洗涤后不能褪色;改进洗涤方法后与对照组均无着色现象.结论 氯己定与次氯酸钠接触可发生化学反应,生成对氯苯胺沉淀,使医用织物着色.     

基于滤膜上细菌直接计数法的细菌总数快速检测

细菌总数快速检测在质量监测中具有重要的意义,目前除了经典的平板培养法以外,还有微菌落法、阻抗法等快速检测方法,这些方法或者需要较长的检测时间,或者需要较高的检测成本.本研究提出一种不需要培养而在滤膜上直接计数的细菌总数的快速检测方法,它主要分为过滤、染色、显微镜计数和计算四个步骤.计算细菌总数时,根据细菌在滤膜上的分布特点,对传统公式进行改进,提出按区域计算细菌总数的计算方法,提高了检测精度.研究结果表明,该方法与传统的平板培养法无显著性差异(t=0.847,P=0.436＞0.05),是一种低成本、快速的细菌总数检测方法.     

Identification d’Enterocytozoon bieneusi par PCR dans les selles des patients immunodéprimés tunisiens

Introduction

Intestinal microsporidiosis is recognised as an important cause of opportunistic parasitosis in immunocompromised patients, especially HIV-infected patients. Enterocytozoon bieneusi is the common causal agent. The diagnosis of intestinal microsporidiosis has usually based on microscopic detection of the spores of microsporidia species in stool samples, requires additional staining techniques as Modified Weber's trichrome stain. However, the detection of the spores can be difficult and species determination, which is important for defining the appropriate treatment, is impossible. Polymerase chain reaction (PCR)-based methods have been successfully used for detection of microsporidian infections. They are more sensitive and are able to identify microsporidia species. The purpose of this study is to identify E. bieneusi to adapt treatment and assess the true prevalence of the intestinal microsporidiosis due to this species in compromised patients in Tunisia.

Patients and methods

One hundred and eighteen stools from immunocompromised patients, with a symptomatology in favour of the intestinal microsporidiosis, were analysed using light microscopy after staining with Modified Weber's trichrome stain and PCR.

Results

Only four were positive by Modified Weber's trichrome stain whereas eleven stools were positive by PCR, giving a prevalence of 20% in HIV-infected patients and 5,35% in human immunodeficiency virus-negative patients.

Conclusion

This study confirms the usefulness of PCR in the diagnosis of the intestinal microsporidiosis due to E. bieneusi. Indeed, PCR has greater sensitivity than Modified Weber's trichrome stain and can identify the species of microsporidia in order to adapt the treatment.     

多种蓝色混合染色在乳腺癌前哨淋巴结活检中的临床应用

目的:探讨多种蓝色染料混合染色鉴别前哨淋巴结(SLN)的成功率及预测乳腺癌腋窝淋巴结转移的准确性;为SLND提供一种较佳的示踪染料。方法:从2005年1月至2006年1月我科收治的乳腺癌病例中选取19例实施了前哨淋巴结活检(snb),分美蓝组(1%美蓝3m l)和复合染色组(1%专利蓝,1%异硫蓝,1%美蓝各1m l混合)两组,于术中在乳腺肿块周围腺体内注射,以鉴别定位SLN,全部病例都实施了腋淋巴结清扫术。结果:SLN鉴别的成功率美蓝组和复合染色组分别为77.8%(7/9)和90%(9/10)(P<0.01);预测腋淋巴结转移状态准确性分别为88.9%(7/8)和100%(10/10)(P>0.05)。结论:与美蓝比较,该混合染料是较理想的乳腺癌SNB的生物活性染料示踪剂。     

青少年生长发育期正常前牙颜色的变化趋势

目的分析上海地区青少年生长发育期间正常前牙颜色的变化趋势。方法选取100名均龄13.2岁的儿童及100名均龄为23.4岁的青少年,分别代表颌面部生长发育高峰的起始与结束年龄。采用日本Shofu公司的Shade-eye自动比配色仪,对2组对象的左上中切牙进行颜色测量和分析。结果从生长发育高峰起始到结束,前牙颜色特性变化趋势如下:(1)色相从以红色为主调向以红色与灰色并存变化(P<0.01);(2)明度存在由明向暗变化(P<0.01);(3)饱和度由浅向深变化(P<0.01);结论在人类生长发育阶段,牙色随年龄而改变,呈一定变化规律,即色相由红到灰、明度由明到暗、饱和度由浅变深的变化趋势。