发电机出口断路器GCB在300MW机组的运用初探

通过分析发电机出口断路器GCB的作用、运行管理、经济性和制造水平,比较论证攀钢发电厂2×300MW CFB煤矸石机组是否采用发电机出口断路器GCB的方案,且重点分析了GCB对300 MW级火电机组电气接线、控制、保护及运行管理的影响,提出了攀钢发电厂300MW级火电机组采用GCB是可行的的观点。     

提高HN700mm×300mm型钢成材率的实践

文章对HN700 mm×300 mm型钢轧制缺陷产生的原因进行了分析,并提出了消除轧制缺陷的方法。通过减少轧制缺陷,提高了HN700 mm×300 mm型钢的成材率,并将消除HN700 mm×300 mm型钢轧制缺陷的经验运用到其它规格H型钢的生产中。     

基于Deform-3D软件对AISI4340钢Φ600mm铸坯开坯工艺参数的模拟数值

利用Deform-3D软件对AISI4340钢Φ600 mm铸坯至300 mm × 300 mm坯的开坯过程工艺参数进行了数值模拟.通过对加热温度、轧制速度、压下率、2道次压下对铸坯心部变形和材料流动影响的研究,分析了开坯成形过程中心部等效应力和材料变形特点,获得了Φ600圆连铸坯开坯成300 mm方坯的成形规律.结果...     

300kA下料异常的分析及处理

通过对300 kA下料异常情况的分析,根据实际生产过程中的处理措施及效果,总结出解决300 kA下料异常现象的方法和措施。     

1500/300破碎机前门框悬挂改造

文章叙述了1500/300破碎机原前门框在使用过程中的缺陷,对前门框频繁更换的原因进行了分析,说明了1500/300破碎机前门框改造的具体方案.     

300kA大型铝电解槽槽壳反变形装置的应用与实践

通过对300 kA大型预焙阳极铝电解槽槽壳变形原因分析,对槽壳在大修冷却过程中变形的防治方法进行了探索,并结合云南铝业300 kA电解槽大修反变形措施应用与实践,着重介绍槽壳反变形装置的研制、应用及效果。     

XWC-300电子电位差计无温度指示值的故障分析及处理

研究了XWC-300电子电位差计与热电偶配合在一起,用于工业生产过程的温度测量、记录和自动控制,分析了XWC-300电子电位差计无温度指示值的故障原因,给出了诊断此故障的方法和修复的步骤,为快速修复XWC-300电子电位差计提供了实用的方法和经验.     

300M离子热扩渗表面强化工艺的研究

利用活性屏离子渗氮(ASPN)技术对300M钢进行快速离子渗氮工艺的研究。分析了工艺参数对渗氮处理后300M的微观组织、显微硬度以及渗层结构的影响。试验结果表明,采用这种新的渗氮工艺可以提高300M的表面硬度,通过控制工艺参数可使表面形成较薄的化合物层或不形成化合物层,达到表面强化的效果。     

立式注塑机预料液缸系统的改进

分析了KT—300型立式注塑机液缸顶料造成塑料螺杆变形的原因,并对液缸结构及液路控制系统进行了改进.     

漆包线浸焊用的抗氧化无铅钎焊料研制

目前使用的无铅焊料品种很多,但使用工作温度大都是300℃以下。超过300℃时,氧化加剧,在浸焊中无法使用,焊料浪费大。为解决高温下(大于300℃)无法正常使用问题,减少材料的氧化,经多次试验、分析,研制出可以在大于300℃以上工作温度浸焊的抗高温无铅软钎焊料,节约了原材料,降低了使用成本,解决了被焊铜线变细的技术问题。     

The usefulness of single-strand DNA conformation polymorphism analysis to detect mutations in protein C deficiency

The standard approach for the molecular genetic analysis of protein C deficiency, polymerase chain reaction (PCR) amplification followed by direct sequencing, although very accurate, is time-consuming. The aim of this study is to investigate the usefulness of a simplified, time-saving screening method for the detection of protein C mutations consisting of the combination of multiplex PCR amplifications using the same primers that were designed for sequencing, followed by single-strand DNA conformation polymorphism (SSCP) electrophoresis analysis performed with one set of conditions. The study was designed in two phases. First, we tested six known point mutations located in different exons of the protein C gene by SSCP. Second, we prospectively studied nine patients with protein C deficiency type I using SSCP as the first screening technique. All the exons were amplified with a common PCR protocol, either as single fragments or as multiplex combinations of several of them. In the retrospective study, three out of the six point mutations were visible as a band shift: 40 T-->G (exon 2), 1432 C-->T (exon 3) and 7253 C-->T (exon 8). In the prospective analysis SSCP detected three different mutations. These mutations were: 6128 T-->C (exon 7), 6216 C-->T (exon 7) and in two probands 8631 C-->T (exon 9). In the five remaining patients we identified only two different mutations by direct sequencing: 6246 G-->A (exon 7) in two patients and 8589 G-->A (exon 9) in four patients. In summary, the results from both studies show that only 60% of all mutations can be detected using this simplified method. It also suggests that a multiple set of conditions, smaller PCR fragments, or both, should be used in order to achieve a sensitivity comparable to sequencing.     

Improved detection of p53 point mutations by dideoxyfingerprinting (ddF)

Two screening techniques for identifying point mutations (single-strand conformational polymorphism (SSCP) and dideoxyfingerprinting (ddF)) were compared to sequencing to determine their efficiency in detecting mutations in exons 5-8 of the p53 tumor suppressor gene. Twelve human glioblastoma cell lines were studied by each of the three methods. Ten mutations were identified by sequencing; of these, 10/10 were detected by ddF, while SSCP detected 6/10 true mutations and falsely identified two presumed mutations not confirmed by sequencing. We examined the impact of parameters which influence DNA conformation (gel temperature, gel composition, and PCR product size) on the ability of SSCP and ddF to detect mutations. The sensitivity of SSCP varied with both gel temperature and the size of the PCR product; in contrast, ddF was not influenced by either gel temperature or product length (up to 460 nucleotides). We conclude that the increased sensitivity of ddF, together with its greater ease of application due to the lack of need for optimization, provides significant advantages over SSCP in screening DNA sequences for the presence of point mutations. Our results also suggest that the incidence of p53 mutations may be underestimated in studies of human cancers which utilize SSCP as the method of mutational screening.     

p53 gene alternation in squamous cell carcinoma of the esophagus detected by PCR-cold SSCP analysis

Mutations of the p53 tumor suppressor gene play an important role in the development of common human malignancies. Previous reports revealed that the frequencies of p53 alternations in esophageal carcinoma varied from 26% to 87%. The clinical significance of p53 alternations is still disputed. In the present study, we used polymerase chain reaction--"cold" single-strand conformation polymorphism (PCR-"cold" SSCP)--to investigate p53 genetic alternations in 63 surgical specimens of esophageal squamous cell carcinoma (ESC). Our experiments showed that the optimum buffer temperature for "cold" SSCP analysis was 14 degrees C while the PCR products were around 200-300 bp in size. Among 63 tumorous samples, p53 alternations was detected in 47 tumors, or an incidence rate of 74.6%. For nontumorous mucosal samples, the incidence of p53 alternations was 55.5% (35/63 samples). Additionally, p53 alternations occurred most frequently at exon 6 (50.8%), followed by exon 7 (33.3%), exon 8 (17.5%) and exon 5 (12.7%). Multiple genetic alternations (> or = 2 exons) between p53 exons 5-8 in the same examined samples were found in 21 (33.3%) of 63 tumors, and in 8 (12.7%) of 63 nontumorous mucosal specimens. Our results further showed that p53 alternations did not correlate with age, depth of tumor invasion, lymph node metastasis, tumor stage, cell differentiation or lymphovascular invasion in ESC (P > 0.05). Moreover, the survival rate in patients with p53 alternations was similar to that in patients without p53 alternations (P > 0.05). In conclusion, PCR-"cold" SSCP is a rapid and sensitive method for identifying p53 genetic alternations. p53 genetic alternations occur with a relatively high incidence for ESC, but p53 abnormality has no impact on prognosis.     

Detection of antineutrophil cytoplasmic antibodies in primary sclerosing cholangitis: a comparison of the alkaline phosphatase and immunofluorescent techniques

SSCP (single-strand conformational polymorphism) is used widely in the field of human biomedicine, but its potential as a population genetics tool for the recovery of nuclear gene genealogies remains to be realized. We describe and illustrate a use for SSCP in the physical isolation of nuclear haplotypes that circumvents several difficulties associated with more conventional cloning procedures. The DNA sequence can be determined directly from the isolated haplotypes and used for phylogenetic inference. SSCP provides a convenient first step toward generating nuclear genealogies for population studies.     

PCR SSCP reveals haplotype related polymorphism of PERB1: a new marker for MHC beta block typing

Many new Major Histocompatibility Complex (MHC) genes have been discovered in the last 5 years. Defining the polymorphism of these new genes may elucidate their function and their relevance to diseases with MHC associations. Polymerase chain reaction and single stranded conformation polymorphism (PCR SSCP) analyses were used to detect sequence polymorphisms of PERB1 demonstrated by comparing the available genomic sequence of four haplotypes. This study showed that PCR SSCP of PERB1 is reproducible. In addition, PERB1 alleles segregate within families together with MHC haplotypes. Typing results from the Forth Asia and Oceania Histocompatibility Workshop (4AOHW) cell panel indicate that the identified polymorphisms of PERB1 are "haplotypic', i.e., unrelated individuals carrying the same MHC ancestral haplotypes carry the same PERB1 SSCP pattern. Interestingly, PERB1 SSCP patterns allow the distinction of ancestral haplotypes which share HLA-B serological specificities, such as HLA-B44 and therefore this analysis can be used to further define MHC haplotypes and thus to improve our understanding of the evolution of this complex.     

Prolonged P300 latency in attention deficit hyperactivity disorder predicts poor response to imipramine

P300 is a cognitive evoked potential that evaluates attention and information processing. This study uses auditory and visual P300 topography to develop a classification of attention deficit hyperactivity disorder (ADHD), and find predictors of treatment response. Of 45 ADHD children ages 6 to 15 treated with pemoline in a previous study, 25 were poor responders. Of these 25, 17 participated in an imipramine treatment protocol. Auditory and visual P300 testing was performed before and after treatment using 31 scalp electrodes. Good and poor responders to imipramine were clinically identical. Poor imipramine responders had longer auditory and visual P300 latencies than good responders. Treatment with imipramine decreased auditory P300 latencies and increased auditory P300 amplitudes. We have previously reported that ADHD patients with small right frontocentral auditory P300 amplitudes respond poorly to pemoline. Thus, P300 topography and latency classifies ADHD into three groups: group 1 with normal P300 topography, and good response to pemoline; group 2 with small right frontocentral auditory P300 amplitudes, poor response to pemoline, and good response to imipramine; and group 3 with long auditory and visual P300 latencies and small right frontocentral auditory P300 amplitudes, and poor response to pemoline and imipramine.     

Single cell detection of beta-thalassaemia mutations using silver stained SSCP analysis: an application for preimplantation diagnosis

Although prenatal diagnosis has reduced the number of beta-thalassaemia births, pregnancy termination is still unacceptable for many couples. Preimplantation genetic diagnosis carried out on the third day after in-vitro fertilization offers an alternative. Here we describe the detection of selected beta-thalassaemia mutations in intron I at the single cell level by the application of nested polymerase chain reaction (PCR) and silver stained single strand conformation polymorphism (SSCP) analysis. A total of 294 single somatic cells of different types was amplified with 96% success and all tested mutations in homozygous and heterozygous form were identified correctly. None of the heterozygous or compound heterozygous samples showed any allele-specific amplification failure after the beta-globin gene amplification from single cells. To assess the efficiency of nested PCR on single blastomeres prior to clinical application, 10 single blastomeres were amplified and gave the expected normal pattern when analysed by SSCP. The main advantage of SSCP, particularly for preimplantation diagnosis, is that it allows the direct visualization of each allele and provides a simple means of assessing allele-specific amplification failure. Our results show that the combination of nested PCR and automated silver stained SSCP analysis offers exceptional resolution, accuracy and speed which are essential for preimplantation diagnosis.     

Charcot-Marie-Tooth disease with intermediate motor nerve conduction velocities: characterization of 14 Cx32 mutations in 35 families

Charcot-Marie-Tooth disease can be inherited either autosomal dominantly or recessively or linked to the X chromosome. X-linked dominant Charcot-Marie-Tooth disease (CMTX) is a sensorimotor peripheral neuropathy in which males have usually more severe clinical symptoms and decreased nerve conduction velocities than do females. CMTX is usually associated with mutations in exon 2 of the connexin 32 (Cx32) gene. DNA from 35 unrelated CMT patients, without the 17p11.2 duplication, but with median nerve conduction between 30 and 40 m/s, were tested for the presence of Cx32 mutations. The entire coding sequence of the Cx32 gene was explored using a rapid nonradioactive technique to detect single-strand conformation polymorphisms (SSCP) on large PCR fragments. Thirteen abnormal SSCP profiles were detected and characterized by sequencing. In addition, systematic sequencing of the entire Cx32 coding region in the remaining index cases revealed another mutation that was not detected by SSCP. A total of 14 mutations were found, five of which were not previously reported. These results demonstrate the high frequency (40%) of mutations in the coding region of the Cx32 gene in CMT patients with intermediate MNCV, without 17p11.2 duplications. Most of these mutations (93%) can be detected by SSCP.