Dystrophin protein and rflp analyis for fetal diagnosis and carrier confirmation of duchenne muscular dystrophy

A pregnant woman with indeterminate Duchenne muscular dystrophy (DMD) carrier status, but with DMD diagnosed in her deceased brother (unavailable for study), presented for prenatal diagnosis, intending to continue the pregnancy only if proven unaffected with DMD with near absolute certainty. Creatine kinase (CK) assays to clarify carrier status were inconclusive. Male sex in the fetus was identified, but DNA restriction fragment length polymorphism (RFLP) analysis was not yet available to this centre to investigate the possible transmission of the DMD gene, and the pregnancy was terminated. Tissue histology and dystrophin protein analysis demonstrated the absence of DMD. In a situation with proven maternal carrier status, future fetal inheritance of the opposite maternal X chromosome would indicate the presence of DMD. However, maternal carrier status remained in doubt through a second pregnancy, even with RFLP studies, and was finally established when dystrophin analysis confirmed the presence of DMD in the second fetus. Histologic findings are presented, contrasting features in the two fetuses. The value of dystrophin analysis for establishing the diagnosis of fetal DMD, in this case proving maternal carrier status in a difficult situation, and for demonstrating DMD gene:RFLP haplotype relationships is illustrated.     

Prenatal diagnosis in a female carrying a deletion close to the duchenne locus

Family studies including the proband are usually needed before a prenatal diagnosis may be performed for Duchenne muscular dystrophy. We report here on prenatal diagnosis in a family where the solitary index case was dead, and where the consultand and her mother were assumed to be carriers by independent evidence. DNA anaylsis revealed that both the consultand and her mother had an X chromosome deleted for DNA material in the Xp21 region. The female fetus also carried the deleted X chromosome.     

Applicability of DNA isolated from syncytiotrophoblast vesicles to gene amplification and molecular analysis

Maternal contamination of fetal DNA represents a major problem when highly sensitive molecular techniques are used in the prenatal diagnosis of genetic diseases. For this reason, we have studied the possibility of using DNA isolated from syncytiotrophoblast vesicles as a target of gene amplification (PCR). Three PCR systems were selected which included a repetitive 149 bp fragment of the Y chromosome, the VNTR locus D1S80, and a portion of the β-globin gene. The results of these experiments indicate that DNA isolated from syncytiotrophoblast vesicles is free of maternal contamination and is suitable for gene amplification and DNA analysis.     

载热体加热炉运行安全探讨

根据载热体(道生油)的性质和加热炉热油系统的特点,结合以往发生的事故进行危险分析,提出道生油加热炉安全操作方法.     

First-trimester prenatal diagnosis of spinal muscular atrophy using microsatellite markers

Twenty-five pregnancies at risk for spinal muscular atrophy I (SMA I) have been monitored by first-trimester prenatal diagnosis. Microsatellite markers were used in all cases to amplify polymorphic regions at the D5S125, D5S435, D5S39, D5S127, and D5S112 loci. All families, including 12 SMA I pedigrees with a deceased index child, were fully informative for DNA analysis. Three fetuses were predicted to be affected and 22 fetuses were predicted to be unaffected. Twenty-two newborns were unaffected by clinical examination at birth. These results support the accuracy of SMA I prenatal diagnosis based on linkage analysis.     

Carrier detection of duchenne muscular dystrophy through analysis of dna from deciduous teeth of a dead affected child

The sister of a child affected by Duchenne muscular dystrophy (DMD) was referred for genetic counselling to assess the risk of her being a carrier. Her brother had died 15 years previously at the age of 8. There were no other affected males in the family. There were no methods for DNA investigation at the time of the child's death and the family had never been studied for linkage with polymorphic probes on the chromosomal region Xp21. The only tissue from which an assessment of the risk could be made by DNA linkage analysis was two of the child's deciduous teeth that the parents had kept. DNA was extracted using a protocol described for the recovery of ancient DNA from museum specimens and archaeological finds. Multiplex amplification did not reveal deletions in 19 exons spanning the hot-spot regions for deletions within the dystrophin gene in Xp21. Linkage analysis using three highly polymorphic microsatellites demonstrated that the sister had not received the X chromosome borne by her brother. These results show that DNA extracted from teeth is a reliable source for molecular diagnosis.     

Lesch—Nyhan syndrome: Carrier and prenatal diagnosis

We report the results of carrier and prenatal diagnosis for hypoxanthine guanine phosphoribosyltransferase (HPRT) deficiency, Lesch—Nyhan syndrome, by carrier testing of 83 women and prenatal analysis of 26 pregnancies. Our diagnostic methodologies include mutation detection and linkage analysis for probands and their families and biochemical measurement of HPRT enzyme activity for at-risk pregnancies. Identification of the mutation in the index case of each family permits precise carrier diagnosis using polymerase chain reaction (PCR) amplification of HPRT gene sequences and automated DNA sequencing. We demonstrate 100 per cent sensitivity for the detection of mutations in the HPRT gene of affected males and highly efficient carrier testing of at-risk females. Two other molecular methods proven to have high utility include PCR-based dosage analysis and linkage analysis by PCR amplification of a short tandem repeat (STR) in intron 3 of the HPRT gene. As a result, 45 at-risk women, 56 per cent of those tested, were identified not to be carriers of their family's HPRT gene mutation. Seven of these women were the mothers of affected males and prenatal testing for future pregnancies was recommended because of the possibility of gonadal mosaicism. Thirty-eight of these women were more distant relatives of affected males, thereby eliminating the need for future prenatal procedures. These studies illustrate the utility and precision of molecular methodologies for carrier and prenatal diagnosis of Lesch—Nyhan syndrome. These studies also illustrate that molecular diagnostic studies of affected males and carrier testing prior to pregnancy can clarify genetic risk predictions and eliminate unnecessary prenatal procedures.     

两种悬浮填料在ASBBR厌氧氨氧化系统中的性能比较与微生物解析

选取聚乙烯（T1）和聚氨酯（T2）两种悬浮填料投入到两套相同厌氧序批式生物膜反应器（ASBBR）R1和R2中,以城市生活污水为配水基质,考察了两种填料在ASBBR中的性能及菌群结构特征,选出更适宜厌氧氨氧化（ANAMMOX）菌生长的填料.结果表明：R1和R2反应器分别在第93 d和73 d成功启动厌氧氨氧化,并实现稳定运行.稳定运行阶段,R1、R2系统总无机氮（TIN）平均去除率分别达88.34%、87.97%.此外,运行第150 d时,测得单个T1、T2填料的ANAMMOX菌活性分别为5.70和3.70 mg·g^-1·h^-1.同时,扫描电子显微镜（SEM）表征结果证明,球状菌在T1填料上的数量比同倍数下T2填料上多,且杂菌较少.高通量测序结果显示,T1、T2填料上Candidatus anammoxoglobus菌属相对丰度分别为75.29%和38.23%.本研究表明,相比于聚氨酯（T2）填料,聚乙烯（T1）填料更适宜ANAMMOX菌的富集.     

Identification of duchenne muscular dystrophy genomic probe P20 constant Taq1 fragment corresponding to the EcoRV and Msp1 polymorphisms

The majority of Duchenne and Becker muscular dystrophy cases are caused by deletions observable in Southern blots with cDNA probes for the gene. When the deletion includes polymorphic probes, they may be used to determine carrier status by deletion segregation analysis: non-inheritance of parental alleles, or heterozygosity. The polymorphic genomic probe P20 is deleted in a large percentage of probands. P20 hybridizes with two constant fragments of 6.7 and 0.8 kb in Taql digests. In a number of probands, only the larger P20 Taq1 fragment is deleted. This study demonstrates that this fragment corresponds with the polymorphic EcoRV and Mspl fragments of P20. Families in which the upper Taql fragment is deleted may be screened for carrier status using non-inheritance of parental alleles or heterozygosity of P20 in EcoRV or Mspl digests.     

基于引物的湖泊沉积物氨氧化细菌PCR扩增策略比较

PCR扩增是检测环境中β-变形杆菌纲氨氧化细菌(β-AOB)群落的主要方法,但是引物的敏感性和特异性对结果具有关键影响.本研究首先比较了2组常用的β-AOB 16S rRNA基因引物,结果显示,在扩增一组不同性质湖泊沉积物样品时,βAMO引物均获得明亮的单一条带,但CTO引物则未能全部扩增.克隆及序列分析证实,βAMO引物扩增获得的序列均不属于β-AOB所在的Nitrosomonadales目,而CTO引物扩增获得的序列来自β-AOB中的Nitrosomonas europaea/\"Nitrosococcus mobilis\"分支.采用变性梯度凝胶电泳方法,对4种不同引物组合PCR策略扩增所得产物进行分析,发现以βAMO或16S rRNA通用引物结合CTO引物的巢式方案可以提高扩增的效率,且β-AOB的群落轮廓与CTO引物直接扩增方案高度相似.这些结果表明βAMO引物具有较高的敏感性但特异性较低,而CTO引物则相反.因此,特定巢式扩增方案既可提高扩增的效率,也能真实反映湖泊沉积物中β-AOB的群落轮廓,是较为理想的β-AOB研究方法.     

富勒烯对PCR反应的抑制作用

以110bp单链DNA为模板,研究富勒烯(C60)对聚合酶链式反应(polymerase chainreaction,PCR)的影响.实验结果表明,随着C60浓度的增加,PCR反应被显著抑制;将Taq DNA聚合酶、单链DNA模板与C60孵育后,其PCR扩增产物均显著减少;增加PCR反应体系中的Taq DNA聚合酶量,可消除C60的抑制作用,但增加起始单链DNA模板的数量,效应不明显.上述研究结果说明,C60不仅可抑制Taq DNA聚合酶活性,同时对DNA模板也具有一定的损伤作用.     

基于新型载体AMC/UASB-SCMBBR生物工艺处理印染废水的中试研究

针对泉州某印染厂实际废水(COD 800～1800 mg·L~(-1),NH~+_4-N 20～50 mg·L~(-1),pH=8～14),以两种新型载体分别作为厌氧与好氧单元中微生物固定化载体,考察了中试规模上流式厌氧污泥床反应器(UASB)与柔性悬浮载体移动床生物膜反应器(SCMBBR)组合工艺对该废水的处理效果.4个月的连续试验结果表明:①经过30 d前期调试运行该工艺系统成功启动,处理量为1.5～2.2 m~3·d~(-1)时,AMC/UASB和SCMBBR反应器对COD的去除率分别达到22%和50%,AMC/UASB反应器出现明显的产气现象,SCMBBR中载体挂膜状况良好;②启动完成后经过90 d的连续运行,当AMC/UASB和SCMBBR反应器水力停留时间分别为10.7 h和8.8 h,废水处理量为5 m~3·d~(-1)时,稳定运行阶段整个工艺废水COD去除率达到78%,出水氨氮平均浓度为3.4 mg·L~(-1);此外,整体工艺色度去除率达到65%,其中,厌氧段色度去除率达到50%左右;③通过分析计算,整个系统污泥减量达到67.7%～76.6%,该生物组合工艺具有明显的污泥减量效果.     

不同填料对减缓MBR膜污染影响的研究

膜生物反应器(MBR)由于存在膜污染问题,使水通过膜的阻力增加,过滤性下降,导致膜通量下降或跨膜压差升高,增加运行费用,限制了其进一步推广应用.通过向MBR中投加填料可以改善膜组件性能、优化系统运行条件以及改善混合液特性,从而减缓膜污染.重点分析了颗粒填料、悬浮填料和絮凝剂的投加对膜污染减缓的影响,进一步展望了可用于改善MBR系统填料的研发价值以及该工艺在污水处理和中水回用领域的良好前景.     

生物膜反应器处理洗涤污水的试验

介绍了采用生物膜反应器处理洗涤污水的试验研究方法，研究了COD、LAS和NH3-N的去除效果。试验结果表明：整个系统出水稳定，水质良好（COD〈38．7mg／L、NH3-N〈1．1mg／L、LAS〈0．9mg／L，且无色无味、无SS），符合国家建设部生活杂用水回用水质标准，且具有较强的抗冲击负荷能力。在生物膜反应器中，微生物对污染物的去除起主要作用，而中空纤维膜对维持系统的稳定出水起重要作用。     

高温ASBR处理热水解污泥的梯度升温法启动试验研究

采用序批式运行方式、梯度升温方法进行了高温ASBR处理热水解污泥的启动实验.结果表明,梯度升温法能在131 d内较快地启动高温ASBR反应器.在3个梯度升温阶段中（35℃→40℃→47℃→53℃）,反应器的性能都出现了一定的波动,其中在中温→中温（35℃→40℃）、高温→高温（47℃→53℃）阶段出现的波动比较小,而在中温→高温（40℃→47℃）阶段反应器的产气量、甲烷含量、出水COD和反应器内微生物量都出现了较大的变化,反应器的性能波动较大,40℃和47℃分别为中温和高温的上限和下限.在反应器启动过程的稳态期,反应器的平均产气量为2.038 L/d,甲烷含量为72.0％,COD产气率（CH₄/COD_投入）为188.8　mL/g,TCOD和SCOD平均去除率分别为63.8％和83.3％.扫描电镜和DGGE分析结果表明,启动过程中反应器中的微生物形态和种类都发生了变化,启动初期（35℃）和稳定期（53℃）的优势菌种明显不同.