Cloning and the identification of major deletion-prone exons of dystrophin gene / 中国组织工程研究

BACKGROUND: Dystrophin gene is X-linkage recessive heredity nerve-muscle system disease. Dystrophin gene deletions cluster in two hotspot regions, comprising exons 2-20 and 44-53. The majority of deletions can be detected by examining only a subset of exons. However, little is known regarding systematic detection of 18 common deletion exons of dystrophin gene.OBJECTIVE: To obtain and identify the cloning of 18 deletion-prone exons of dystrophin gene.METHODS: A total of 18 fragments of dystrophin gene were obtained through polymerase chain reaction (PCR) amplification with human genomic DNA as template and 18 pairs of primers respectively. The fragments were connected with pGEM-T Easy vector.The recombinants were transformed into E.coli JM109 competent cells, followed by planted on Luria-Bertani (LB)/ampicillin(Amp)/isopropylthio-β-D-galactoside(IPTG)/X-bromo-4-chloro-3-indolyl-β-D-galactoside (X-Gal) plates and cultured.Positive transformants were selected with blue/white color screening, and the recombinant plasmids DNA was extracted and digested with restriction enzyme Not I. DNA sequences of the fragments were analyzed. Nucleotide analyses were performed through the National Center for Biotechnology Information (NCBI) Basic Local Alighment Search Tool (BLAST) against GenBank.RESULTS AND CONCLUSION: Size of the18 fragments by PCR amplification was in accordance with anticipation. Size of the fragments of recombinant cloning by Not I digestion was in accordance with that of PCR and expectation. Sequence size of the 18cloned fragments was in accordance with expectation. The cloned fragments have high homology with dystrophin gene through NCBI BLAST against GenBank. These cloned fragments were the main deletion-prone exons of dystrophin gene.

Guillain-Barré syndrome in northwestern China / 中国组织工程研究

We reviewed 100 cases with Guillain-Barré syndrome (GBS) from 1980 to 1999, and found that the features of GBS in electrophysiological classification, age, area, seasonal distributions, and in preceding illness in northwestern China are different in some aspects from those in Europe and North America or in northern China. The demyelinating pattern appeares as a major subtype not only in different age groups, but also in different test times after symptom onset.

Electrophysiological classification of Guillain-Barr? syndrome: analysis of 100 cases / 中华神经科杂志

Objective To explore the electrophysiological and clinical features of various subtypes of Guillain Barr? syndrome (GBS). Methods The electrophysiological and clinical data of 100 cases with GBS admitted to Xijing Hospital from 1980 to 1999 were analyzed retrospectively. Correlations between varied subtypes and ages were examined by ? 2 test. Results Among the 100 patients with GBS, the demyelinating pattern was present in 51 patients, the axonal pattern in 25 patients, and 8 patients were inexcitable, 12 patients equivocal and 4 patients normal. The demyelinating pattern appeared as a major subtype not only in different age groups, but also in different test times after symptom onset. There was no statistically significant relationship between varied subtypes and ages. In the 100 patients, 32.0% suffered from a preceding upper respiratory infection, and 22.0% had a preceding gastrointestinal tract infection. The cases occurring in rural areas are almost in number equal to those in urban areas. That is, there was no a clear area distribution. Both demylinating and axonal GBS occurred throughout the year with a likely peak from July to September. Conclusion In the 100 patients with GBS admitted to Xijing Hospital, the demyelinating pattern was the major electrophysiological subtype. In addition, the electrophysiological and clinical features of various subtypes of GBS seemed to be different in some ways from those in the studies of both western countries and Li CY in northern China.

Detection of Duchenne and Becker muscular dystrophy patients by DNA microarray / 临床神经病学杂志

Objective To explore the efficient method in detection of DMD/BMD patients.Methods 18 deletion-prone exon fragments of DMD gene were amplified via molecular cloning. They were used as probes and were spotted on the slides treated with APES and poly-lysine together by manual operation to make microarray. In addition, fragments of ?-actin were used as positive contrast and those of pUC 19/EcoR I were used as negative. 30 DMD/BMD patients were detected for deletion in DMD gene with the microarray and 5 healthy people were done as normal control. Parts of the results were compared with PCR method.Results Different exon fragment deletion of DMD/BMD gene was detected in 21 patients by DNA microarray, and 10 of them were confirmed by PCR analysis.Conclusion DNA microarray assay is a convenient ,accurate and sensitive method in diagnosis of DMD/BMD patient.