Eleven novel mutations in the factor VIII gene from Brazilian hemophilia A patients

The molecular characterization of the mutations in hemophilia A patients is hampered by the large size of the factor VIII gene and the great heterogeneity of mutations. In this study, we have performed a protocol involving multiplex polymerase chain reaction in which 19 exons were amplified in four different combinations followed by nonradioactive single-strand conformational polymorphism (SSCP) to screen for mutations. Southern blotting was used to detect inversion of the factor VIII gene resulting from recombination between copies of the gene A (F8A) located in intron 22 of the factor VIII gene and two copies close telomeric region of X chromosome. Forty-two hemophilia A patients (21 with severe and 21 with mild-to-moderate disease) were studied. The inversion of factor VIII occurred in 13 of 21 patients affected by severe hemophilia A. One patient showed a large extra band in addition to the three bands observed after Southern blotting with the F8A probe. An abnormal electrophoretic pattern of SSCP was detected in 85% and 50% of the patients affected by mild-to-moderate and severe disease, respectively. Sixteen different mutations were identified. Eleven mutations were novel and comprised 9 point mutations and 2 small deletions. This study shows that the methodology used is safe and rapid and has potential for detecting almost all of the genetic defects of the studied hemophilia A patients.     

Detection of the hemoglobin E mutation using the color complementation assay: application to complex genotyping

The color complementation assay (CCA) is a method of allele-specific DNA amplification by which competitive priming and extension of fluorescently labeled oligonucleotide primers determine the color of DNA amplification product. This diagnostic method precludes the need for radioisotopes, electrophoresis, and multiple high-stringency reaction conditions. The multiplicity of mutant globin genes present in Southeast Asians complicates clinical diagnosis and underscores the importance of DNA-based diagnostic methods. We have applied CCA to distinguish beta A and beta E alleles. Competing 15mer primers were a fluorescein-labeled complement to beta A and a rhodamine-labeled complement to beta E, identical except for their central nucleotides. A common unlabeled primer was used to amplify DNA product, the color of which was determined by the perfectly complementary primer. Color photography and spectrofluorometry, as well as a method of black-white photography that we developed to distinguish fluorescein- and rhodamine- labeled DNA, were used to record results. We applied CCA to define the complex genotype of a Thai woman with thalassemia intermedia, 96% HbE, and 4% HbF whose possible genotypes included several permutations of alpha-thalassemia, beta-thalassemia, and beta E genes. zeta-Globin gene mapping of DNA doubly digested with Bg/II and Asp 718 showed the -alpha 3.7/--SEA genotype, and CCA confirmed homozygous beta E/beta E. The CCA is useful for diagnosing the compound hemoglobin genotypes of Southeast Asians and could be applied also to prenatal diagnosis in this population.     

Effects of thick Tenon's capsule on primary trabeculectomy with mitomycin-C

PURPOSE: To investigate the effects of thick Tenon's capsule on primary trabeculectomy with adjunctive mitomycin-C. METHODS: In this prospective interventional case series of 45 consecutive uncomplicated glaucoma patients, 45 eyes with thick Tenon's capsule underwent primary trabeculectomy with intraoperative mitomycin-C (0.4 mg/ml for 3 min). Success was defined as intraocular pressure (IOP) 相似文献   

Laryngeal atresia presenting as fetal ascites, olygohydramnios and lung appearance mimicking cystic adenomatoid malformation in a 25-week-old fetus with Fraser syndrome.

We describe a 25-week-old female fetus of consanguineous parents with ultrasonographic findings of increased echogenicity of lungs mimicking CAM (cystic adenomatoid malformation) type III, olygohydramnios and fetal ascites. A therapeutic abortion was performed and unilateral cryptophthalmos, laryngeal atresia and bilateral syndactyly of the hands and feet were observed at post-mortem. These findings confirmed the diagnosis of Fraser syndrome after abortion.     

垂全催乳素瘤的诊断和治疗指南

3.男性:高催乳素血症通常导致阳痿、不孕和性功能低下.男性患者通常为大腺瘤,有神经系统症状.其原因可能是对症状认识的延误或者肿瘤生物学行为的差异.     

Lentiginous macules and patches of neurofibromatosis (an approach to better terminology)

BACKGROUND AND OBJECTIVES: Freckles (ephelides) are small, light brown macules of sun-exposed areas in fair-skinned subjects. On the other hand, freckle-like pigmentation of the axilla is a highly characteristic feature of neurofibromatosis. To what extent (clinically and pathologically) are the two pigmentary defects similar? And to what extent are café-au-lait patches and freckle-like lesions similar pathologically? SUBJECTS AND METHODS: Ten cases of neurofibromatosis and 10 cases of freckles were examined clinically. Two biopsies were taken from the former; one obtained from an axillary freckle-like lesion, and the other from a café-au-lait patch. One biopsy only was taken from freckles. The biopsies were processed for haematoxylin and eosin, silver stain and transmission electron microscopy. RESULTS: Obvious differences were detected between freckles (ephelides) and freckle-like lesions in cases of neurofibromatosis. Café-au-lait patches and freckle-like lesions showed the same pathological changes of lentigo simplex, with increased melanin pigment from the basal layer up to the stratum corneum, moderate elongation of the rete ridges and mild inflammatory infiltrate intermingled with melanophages. Electron microscopy revealed an increase in melanocyte number and the presence of giant melanin granules in freckle-like pigmentation as well as café-au-lait patches. These ultrastructural freckles (ephelides) showed an increase in activity but not the number of melanocytes. CONCLUSIONS: The term freckle-like should be changed to lentiginous macules of neurofibromatosis. On the other hand, the café-au-lait patches should be considered as lentiginous patches. Freckles should be restricted to sun-exposed areas in fair-skinned persons only.     

接受肾移植前维持性血液透析患者生活质量与静脉补充左旋卡尼汀的影响

目的:血液透析患者大多存在卡尼汀尤其是游离卡尼汀的缺乏,透析后补充左旋卡尼汀能够减轻、减少并发症及住院率。观察静脉补充左旋卡尼汀对肾移植前血液透析患者生活质量的影响。方法:选择2005-09/2006-09于柳州市人民医院进行维持性血液透析的患者30例,对治疗方案均知情同意。按随机数字表法分为左旋卡尼汀组和对照组,每组15例。每次透析结束后,左旋卡尼汀组静脉注射左旋卡尼汀20mg/kg,对照组注射等量的生理盐水,用药8周。用药前及用药8周时分别应用SF-36量表进行生活质量评分,由8个方面组成,每项都以0￣100分计,得分越高表示生活质量越好;同时记录透析相关症状及实验室参数,比较两组间差异。结果:30例患者全部进入结果分析。①与用药前相比,用药8周后左旋卡尼汀组SF-36总体评分增加了(18.29±12.71)分,对照组总评分减少了(6.40±16.39)分。与对照组相比,左旋卡尼汀组总评分及生理功能、总体健康、活力、社会功能、精神健康评分均显著增加(P<0.05￣0.01)。②与对照组相比,左旋卡尼汀组血红蛋白、白蛋白含量均显著增加(P<0.01)。结论:静脉补充左旋卡尼汀不仅有助于改善维持性血液透析患者的贫血状况,而且能明显提高其生活质量。