联合应用二维及三维超声对胎儿脑膨出的诊断价值

目的 探讨联合应用二维及三维超声对胎儿脑膨出的诊断价值.方法 分析6例经引产后证实为胎儿脑膨出患者二维及三维超声表现.结果 二维及三维超声能清晰显示胎儿颅骨缺损的部位、大小,根据膨出物的特点判断脑膨出的类别.结论 联合应用二维及三维超声检查在诊断胎儿脑膨出畸形中具有重要的诊断价值.     

2D-PAGE analysis of Taenia solium metacestode 10-30 kDa antigens for the serodiagnosis of neurocysticercosis in children

Neurocysticercosis (NCC) caused by T. solium metacestode is an increasingly important health issue in Indian children. The sensitivity and specificity of available serological techniques were low in case of single cysticercus granuloma cases which is a more common feature in Indian patients who are children. Serum samples were collected from 13 clinically and radiologically suggestive NCC children and seropositive by ELISA, 25 clinically and radiologically suggestive NCC children and seronegative by ELISA and 25 control subjects. The 10–30 kDa antigens of T. solium metacestode were subjected to 2-dimensional gel electrophoresis (2D-PAGE) followed by enzyme-linked immunoelectrotransfer blot (EITB) assay to detect antibody in serum. Analysis of 10–30 kDa antigenic fraction 2D-PAGE map showed 31 proteins between 10 and ≤28 kDa and innumerable proteins between >28 and 30 kDa with the Isoelectric point of 3–10. All the 13 (100%) NCC seropositive and 15 (60%) out of 25 NCC seronegative samples were reactive with 2D fraction antigens. In the control group, none of the serum was reactive except 2 hydatid samples (92% specificity). The sensitivity and specificity of 2D-PAGE EITB assay were significantly higher than the ELISA which is the routine diagnostic method used in the endemic countries for the serodiagnosis of neurocysticercosis.     

Integrated sample cleanup and capillary array electrophoresis microchip for forensic short tandem repeat analysis

A twelve-lane capillary array electrophoresis (CAE) microsystem is developed that utilizes an efficient inline capture injection process together with the classical radial microfabricated capillary array electrophoresis (μCAE) format for high-sensitivity forensic short tandem repeat (STR) analysis. Biotin-labeled 9-plex STR amplicons are captured in a photopolymerized gel plug via the strong binding of streptavidin and biotin, followed by efficient washing and thermal release for CE separation. The analysis of 12 STR samples is completed in 30 min without any manual process intervention. A comparison between capture inline injection and conventional cross injection demonstrated at least 10-fold improvement in sensitivity. The limit-of-detection of the capture-CAE system was determined to be 35 haploid copies (17–18 diploid copies) of input DNA; this detection limit approaches the theoretical limits calculated using Poisson statistics and the spectral sensitivity of the instrument. To evaluate the capability of this microsystem for low-copy-number (LCN) analysis, three touch evidence samples recovered from unfired bullet cartridges in a pistol submerged in water for an hour were successfully analyzed, providing 53, 71, and 59% of the DNA profile. The high-throughput capture-CAE microsystem presented here provides a more robust and more sensitive platform for conventional as well as LCN and degraded DNA analysis.     

聚丙烯酰胺凝胶垂直电泳和琼脂糖水平电泳检测基因型的比较研究

目的比较聚丙烯酰胺凝胶电泳（polyacrylamide gel electrophoresis,PAGE）垂直板法-银染和琼脂糖凝胶水平电泳应用于环氧合酶-2（cyclooxygenase-2,COX-2）-1195GA基因型检测的效果。方法由两名实验人员分别应用PAGE垂直板法-银染和琼脂糖水平电泳两种方法读取100例患者的COX-2-1195GA基因型,比较结果一致性以及两种方法的特点。结果两种方法检测结果符合率100%。PAGE垂直板法-银染灵敏度较高,但技术难度较大,操作时间长,成本高,图片较难编辑;琼脂糖凝胶水平电泳可分离相差50 bp的片段,且操作简单,节省时间,图片易编辑。结论在酶切片段易于分离检测时,琼脂糖凝胶水平电泳可推广应用于聚合酶链反应—限制性片段长度多态性分析的基因型检测。     

特发性高草酸尿症大鼠空肠蛋白质组二维电泳可行性研究

目的：在正式进行双向凝胶电泳实验前需要进行二维电泳的可行性评价,为双向凝胶电泳实验的顺利进行奠定实验基础。方法：取1只特发性高草酸尿大鼠（IH）和正常对照大鼠的空肠组织300mg,匀浆提取组织总蛋白,以双向电泳技术分离蛋白质,银染后扫描获得图谱,并以ImageMaster^TM 2D Platinum software（Version5．0）软件进行分析。结果：获得了清晰、稳定的凝胶蛋白图谱,正常大鼠凝胶图谱可检测到2541个蛋白点,IH大鼠凝胶图谱可检测到2654个蛋白点。结论：从获得的二维电泳图谱来看,2个蛋白质样品的蛋白质点的迁移基本稳定,从而保证后续正式的双向凝胶电泳实验有较好的重复稳定性和可行性。     

The diagnostic dilemma of congenital unstable hemoglobinopathies

Unstable hemoglobin variants represent a rare etiology of congenital hemolytic anemia. Without a high index of suspicion, plus proper laboratory testing and interpretation, the correct diagnosis can be elusive. We report on five children who were initially thought to have other congenital disorders such as hereditary spherocytosis or thalassemia, before β‐globin gene sequencing led to the definitive diagnosis. Recognizing the variable clinical presentation and laboratory data reported will aid clinicians in diagnosis of unstable hemoglobins variants in children with atypical forms of hemolytic anemia, particularly those with low pulse oximetry values or whose hemoglobin electrophoresis suggest β‐thalassemia trait. Pediatr Blood Cancer. 2010;55:1393–1395. © 2010 Wiley‐Liss, Inc.     

A simplified method of determining synovial fluid chondroitin sulfate chain length

OBJECTIVE: To determine whether dimethylmethylene blue (DMMB) analysis, when combined with agarose gel filtration chromatography (Superose 6), can be performed instead of fluorophore-assisted carbohydrate electrophoresis (FACE) to determine chondroitin sulfate (CS) chain length in synovial fluid (SF). METHODS: SF was obtained from (1) normal horses after 8 weeks of rest, (2) the same horses after 9 months of treadmill training, and (3) horses with osteochondral (OC) injury from racing. SF CS concentrations and chain lengths were determined by gel chromatography and DMMB analysis and compared with previous results determined by FACE analysis on the same samples. RESULTS: DMMB analysis showed that SF CS peak chain length in the OC injury group increased significantly (18.7 kDa) when compared to rested and exercised normal horses (15.6 kDa). The assay had a positive predictive value of 71% and a negative predictive value of 75% for discriminating between normal and injured joints. CONCLUSIONS: We report a simple and inexpensive DMMB analysis of SF CS chain length, which, when coupled with Superose 6 chromatography, discriminates between normal and post-injury joints. Similar to our previous FACE analysis results [Brown MP, Trumble TN, Plaas AHK, Sandy JD, Romano M, Hernandez J, et-al. Exercise and injury increase chondroitin sulfate chain length and decrease hyaluronan chain length in synovial fluid. Osteoarthritis Cartilage 2007;15], our DMMB results show an increase in the chain length of the CS in the SF of injured joints.     

Simultaneous determination of streptomycin and oxytetracycline in agricultural antimicrobials by CZE after an experimental design

A capillary zone electrophoresis (CZE) method was developed and validated for the simultaneous determination of both streptomycin (STP) and oxytetracycline (OTC) in bactericidal products to be used in agriculture. Using fused-silica capillaries, the influence of the electrolyte composition, pH and concentration, as well as temperature and applied voltage were investigated using a central composite design to optimize the method. The optimized electrophoretic conditions were as follows: 0.10 M sodium phosphate, pH 2.5, 7.0 kV and 20.0 degrees C. The method was validated for STP and OTC determination in agricultural formulations through the following performance criteria: linearity and linear range, sensitivity, selectivity, intra-day and inter-day precision, detectability, accuracy and ruggedness. This optimized CZE-method for the identification and quantification of STP and OTC is a potential alternative method to the HPLC methods described by the US Pharmacopeia, with the advantage that the same method could be used for the simultaneous determination of these different antibiotics.     

Determination of kava lactones and flavonoid glycoside in Scorzonera austriaca by capillary zone electrophoresis

A capillary zone electrophoretic method has been developed for the quantitative analysis of three active comppounds, 12-hydroxy-desmethoxyyangonin (HD), 12-beta-d-glucopyranoside-desmethoxyyangonin (GD) and luteolin 3'-(6-E-p-coumaroyl-beta-d-glucopyranoside) (LG) in Scorzonera austriaca with UV detection at 254 nm. The applied voltage was 25 kV and the capillary temperature was kept constant at 25 degrees C. The effect of buffer pH, the concentration of electrolyte and organic modifier on migration were studied systematically. Optimum separation was achieved with 20 mM borate buffer at pH 10.00 containing 10% (v/v) methanol. Daphnetin was used as internal standard for quantification. Regression equations revealed good linear relationship between the ratios of the peak area of each compound and its the ratios of concentration. All the correlation coefficients were higher than 0.9990. The relative standard deviations of migration time and the peak area were <1.46% and 5.13% (inter-day), and <1.65% and 5.16% (intra-day), respectively. The contents of the three compounds in S. austriaca were successfully determined with satisfactory repeatability and recovery.